DE112013002484T5 - Pteridine ketone derivative and uses thereof as EGFR, BLK and FLT3 inhibitor - Google Patents

Abstract

Provided are a pteridine ketone derivative used as EGFR, BLK and FLT3 inhibitor, and applications thereof. In particular, there are provided a compound of the following formula I, a pharmaceutical composition containing the compound of formula I and the use of the compound in the manufacture of a medicament for the treatment of diseases mediated by EGFR, BLK or FLT3 or the EGFR, BLK and FLT3 inhibit.

Description

Area of Expertise

The present invention relates to the synthesis of pteridine ketone compounds and uses thereof in the field of medicinal chemistry and pharmacotherapy. In particular, the invention relates to the use of pteridine ketone compounds having various substituents as EGFR, BLK and FLT3 inhibitors, in particular the use in the manufacture of medicaments for the treatment of tumor-related diseases.

background

A malignant tumor is a cell disorder characterized by out-of-control normal cell division leading to excessive cell differentiation and proliferation, invasion into local tissues, and metastasis. Malignant tumors have become common diseases that pose a serious threat to human life and health. According to the incomplete statistical data, there are about 20 million new cases a year in the world. Therefore, it is currently the most challenging and important area of the life sciences to explore and develop antitumour agents.

In general, conventional antitumor agents are cytotoxic agents that have disadvantages such as unavoidable toxic side effects, low selectivity and resistance, etc. Various basic life events in malignant tumor cells, such as signal transduction, cell cycle regulation, angiogenesis and the like, have become more gradual with the rapid progress of the life sciences demonstrated. It is an important aspect in the study of antitumor agents to develop antitumor agents with excellent therapeutic efficacy and low side effects by using certain key enzymes that are important for the propagation of tumor cells on signal transduction pathways as targets for drug screening. Protein tyrosine kinase is a type of protein for catalyzing the transfer of γ-phosphate from ATP to a particular amino acid residue of a protein that plays an important role in the intracellular signaling pathway and regulates a number of physiological processes such as cell growth, differentiation and death , According to the information in the prior art, more than 50% of the oncogenes and their products possess protein tyrosine kinase activities whose aberrant expression leads to disruption of the life cycle of cells and thereby to the formation of a tumor. Moreover, the aberrant expression of tyrosine kinase is closely related to the metastasis and chemoresistance of tumors.

Epidermal growth factor receptor tyrosine kinase (EGFR) can modulate multiple signaling pathways, direct the extracellular signal into the cell and play an important role in regulating the proliferation, differentiation and apoptosis of normal cells and tumor cells (Cell, 2000, 100, 113-127 ). Therefore, the purpose of treating tumors can be achieved by selectively inhibiting the EGFR-modulated signal transduction pathway, which opens a viable pathway for the targeted treatment of tumors. The active ingredients with EGFR as a target, such as gefitinib, erlotinib and laptinib, are marketed to treat non-small cell lung cancer and breast cancer. However, clinical experience has shown that most patients with non-small cell lung cancer become resistant after being treated repeatedly using gefitinib or erlotinib, with 50% of resistance cases being relevant for the mutation of a single amino acid in the EGFR kinase domain (Threonine residue at position 790 mutated to methionine, T790M) (The New England Journal of Medicine, 2005, 352, 786-792). To overcome T790M-related resistance, a number of irreversible ATP-competitive inhibitors, such as CI-1033, BIBW2992, HKI-272, PF00299804, etc., have been clinically investigated. The irreversible inhibitors have a single Michael receptor fragment that can covalently bind to a single conservative amino acid (Cys797) in the ATP binding center of the EGFR, providing greater EGFR binding affinity than a reversible inhibitor (Journal of Medicinal Chemistry, 2009, 52, 1231-1246). The results of clinical experiments for the above-mentioned irreversible inhibitors are not ideal because of toxicity from off-target effects, low selectivity side effects, and inadequate drug concentration in a patient in vivo, etc. (Nature, 2009, 462, 1070-1074) , Therefore, the development of new irreversible EGFR inhibitors has great clinical importance and good application prospects.

B lymphocyte tyrosine kinase (BLK) belongs to the group of non-receptor tyrosine kinases and is classified as c-Src, Fyn, Lck, c-Yes, Fgr, Hck, Lyn, etc. in the Src family. BLK is mainly used in the B lymphocytes line is expressed, and during the entire process of development of B lymphocytes except the plasma cell phase BLK is expressed. BLK participates in downstream signaling of the B lymphocyte receptor (BCR) (Molecular Biology Reports, 2011, 38, 4445-4453) and affects pre-B lymphocyte receptor-related functions (Journal of Experimental Medicine, 2003, 198 , 1863-1873) and thereby influences the differentiation and proliferation of B lymphocytes. Constitutively activated BLK expressed in murine B-cell and T-cell lines results in the development of B-cell lymphoma or T-cell lymphoma (Proceedings of the National Academy of Sciences, 1998, 95, 7351 -7356). More importantly, ectopic expression of BLK is present in human cutaneous T-cell lymphoma (CTCL) (Blood, 2009, 113, 5896-5904), suggesting that BLK may be used as a potential target for an anticancer drug , In addition, the polymorphism of the BLK gene is closely related to the pathogenesis of systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and other autoimmune diseases (The New England Journal of Medicine, 2008, 358, 900-909), and the induction of a B cell cell apoptosis may be an effective treatment for the above disorders (Nat Reviews Immunology, 2006, 6, 394-403).

FMS-type tyrosine kinase 3 (FLT3), which belongs to the family of type III receptor tyrosine kinases, plays an important role in the proliferation, differentiation and apoptosis of hematopoietic cells (Oncogene, 1993, 8, 815-822). Upon binding to an FLT3 ligand, FLT3 activates several downstream signaling pathways including the STAT5, Ras / MAPK, and PI3K / AKT pathways. FLT3 mutations occur in about one-third of patients with acute myeloid leukemia (AML) (Blood, 2002, 100, 1532-1542), including mutations in internal tandem duplication sequences of the juxtamembrandomain of exons 14 and (or) 15 (FLT3-ITD) and mutations in the sense of an amino acid deletion or insertion in the activation loop of the tyrosine kinase domain (FLT3-TKD). In addition, high expression of FLT3 is found in cases of acute leukemia (Blood, 2004, 103, 1901), and overexpression of FLT3, the FLT3-ITD mutation, and the FLT3-TKD mutation results in poor AML patients Forecast. FLT3 has thus become an important target for the treatment of AML. So far, no FLT3 inhibitors have yet been approved for clinical use, and the clinical effects of many FLT3 inhibitors in clinical trials are still unsatisfactory.

Therefore, one focus of research and development of targeted anti-tumor agents is to improve the clinical efficacy of small molecule kinase inhibitors, and the most promising strategy is to develop multi-target inhibitors containing several kinases associated with the pathogenesis of a disease ( a tumor) are connected simultaneously.

Brief description of the invention

The inventors establish a virtual screening platform for EGFR, BLK, FLT3-specific small molecule inhibitors using computer-aided drug design. Taking full account of the pharmacophore and molecule docking procedures, the inventors searched commercial compound databases (including ACD-3D (chemical library), ACD-SC, MDDR-3D (drug activity data library) and CNPD) and found a panel of potential EGFR candidates. , BLK, FLT3 inhibitory activity.

The structure of the obtained candidate compounds was optimized, and a series of hitherto undescribed pteridine ketone compounds were designed and synthesized, the structure of which was characterized. The activities of the compounds were tested at the molecular and cellular levels, yielding compounds with high EGFR-inhibiting activity. Among these, Compound 032 has IC ₅₀ values of 3.67, 2.36 and 1.17 nM for the inhibition of the activities of the EGFR ^WT , EGFR ^L858R and EGFR ^{T790M / L858R kinases, respectively,} and 0.004 and 0.038 μM for the inhibition of the Propagation of HCC827 cells (non-small cell lung cancer cell, EGFR ^{del E746-A750} ) or, H1975 cells (non-small cell lung cancer cell, EGFR ^{L858R / T790M} )

The pteridine ketone compounds according to the present invention can be used as EGFR inhibitors to block the phosphorylation of EGFR and to inhibit the growth, proliferation and differentiation of tumor cells and therefore these compounds can be developed into novel antitumor agents. In addition, the pteridinketone compounds of the present invention possess high B lymphocyte kinase (BLK) and FMS type tyrosine kinase 3 (FLT3) inhibitory activities and can be used in the development of drugs for the treatment of tumors and immune disorders.

wobei
A und B Benzolringe oder fünf- oder sechsgliedrige Heterocyclen mit verschiedenen Substituenten sind;
C aus einer der folgenden Gruppen ausgewählt ist:

wobei X aus O, S und Se ausgewählt ist, R¹ Wasserstoff, ein Halogenatom, C₁-C₆-Alkoxy (z. B. Methoxy, Ethoxy usw.), ein gegebenenfalls substituiertes C₁-C₆-Alkyl (z. B. ein halogensubstituiertes Alkyl), ein gegebenenfalls substituiertes Aryl (z. B. ein halogensubstituiertes Aryl) oder ein gegebenenfalls substituiertes Aralkyl (z. B. ein Arylmethyl) ist;
R² jeweils unabhängig aus Wasserstoff, einem Halogen, einem C₁-C₆-Alkoxy, Hydroxy, gegebenenfalls substituierten Acyloxy, Amino, gegebenenfalls substituierten Acylamino, gegebenenfalls substituierten C₁-C₆-Alkyl, CN, einer Sulfonatgruppe, Sulfamoyl, Carbamoyl, Carboxy, gegebenenfalls substituierten Alkoxyformyl, gegebenenfalls substituierten Phenyl, gegebenenfalls substituierten N-Alkylpiperazinyl, gegebenenfalls substituierten Morpholinyl, gegebenenfalls substituierten Piperidinyl, gegebenenfalls substituierten Pyrrolyl, gegebenenfalls substituierten Pyrrolidinyl, -NR_aR_b, gegebenenfalls substituierten Pyridyl ausgewählt ist;
R³ jeweils unabhängig aus Wasserstoff, einem Halogen, einem C₁-C₆-Alkoxy, Hydroxy, gegebenenfalls substituierten Acyloxy, Amino, gegebenenfalls substituierten, Acylamino, gegebenenfalls substituierten C₁-C₆-Alkyl, CN, einer Sulfonatgruppe, Sulfamoyl, Carbamoyl, Carboxy, gegebenenfalls substituierten Alkoxyformyl, gegebenenfalls substituierten Phenyl, gegebenenfalls substituierten N-Alkylpiperazinyl, gegebenenfalls substituierten Morpholinyl, gegebenenfalls substituierten Piperidinyl, gegebenenfalls substituierten Pyrrolyl, gegebenenfalls substituierten Pyrrolidinyl, -NR_aR_b, gegebenenfalls substituierten Pyridyl ausgewählt ist;
R_a und R_b unabhängig aus einem Alkyl und einem Alkenyl ausgewählt sind; und
m und n unabhängig aus 0, 1, 2, 3 oder 4 ausgewählt sind.The pteridine ketone compounds according to the present invention have the structure of general formula I:

in which
A and B are benzene rings or five- or six-membered heterocycles with different substituents;
C is selected from one of the following groups:

wherein X is selected from O, S and Se, R ^{1 is} hydrogen, a halogen atom, C ₁ -C ₆ alkoxy (e.g., methoxy, ethoxy, etc.), an optionally substituted C ₁ -C ₆ alkyl (e.g. A halo-substituted alkyl), an optionally substituted aryl (e.g., a halo-substituted aryl), or an optionally substituted aralkyl (e.g., an arylmethyl);
R ^{2 are} each independently selected from hydrogen, halogen, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, sulfonate group, sulfamoyl, carbamoyl, Carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl;
R ^{3 are} each independently selected from hydrogen, a halogen, a C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted, acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, a sulfonate group, sulfamoyl, carbamoyl , Carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl;
R _a and R _{b are} independently selected from an alkyl and an alkenyl; and
m and n are independently selected from 0, 1, 2, 3 or 4.

In one embodiment, R ^{3 is} independently selected from hydrogen, hydroxy, an optionally substituted acyloxy, an optionally substituted amino, an optionally substituted acylamino, an optionally substituted C ₁ -C ₆ alkyl, CN, a sulfonate group, sulfamoyl, carboxy, and optionally substituted Alkoxyformyl selected.

In one embodiment, C is a group of the following formula:

In one embodiment, R ^{1 is selected} from H and an alkyl.

In one embodiment, both A and B are optionally substituted phenyl.

In one embodiment, R ^{2 is} independently selected from H, alkoxy, morpholinyl, halo, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b , acylamino and carbamoyl (NH ₂ C (O) -), where R _a and R _b are selected from alkyl and alkenyl.

In one embodiment, R ^{2 is} independently selected from 4-N-methylpiperazinyl, N-morpholinyl, N-piperidinyl, N-pyrrolyl, N-pyrrolidinyl, N, N -diethylamino, an N, N-dimethylmethylamine group and 4-pyridyl.

In one embodiment, R ^{3 is} independently selected from hydrogen, amino, acyloxy, alkoxy, halo, hydroxy, alkyl, CN, a sulfonate group, sulfamoyl, carboxy, morpholinyl, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b , Acylamino and carbamoyl, wherein R _a and R _b are selected from alkyl and alkenyl.

In one embodiment, R ^{3 is} independently selected from acylamino, acyloxy and alkoxy.

wobei X ein Halogen ist.In one embodiment, R ^{3 is} independently selected from the following groups:

where X is a halogen.

In one embodiment, R ^{3 is} independently selected from the following groups:

In one embodiment, R ^{3 is selected} from the following groups:

In one embodiment, m = 1 or 2.

In one embodiment, n = 1, 2, 3 or 4.

In one embodiment, in group C, the wavy bond in the structural unit comprising R ^{1 is} attached to C, while the other part is bound to NH.

wobei
Y aus N, CH ausgewählt ist;
Z aus N, CR⁶ ausgewählt ist;
R¹ Wasserstoff, Halogen, C₁-C₆-Alkoxy, ein gegebenenfalls substituiertes C₁-C₆-Alkyl, ein gegebenenfalls substituiertes Aryl oder ein gegebenenfalls substituiertes Aralkyl ist;
R³ unabhängig aus Wasserstoff, Amino, Hydroxy, gegebenenfalls substituierten Acyloxy, Alkoxy, Halogen, gegebenenfalls substituierten Alkyl, CN, einer Sulfonatgruppe, Sulfamoyl, Carboxy, Morpholinyl, N-Alkylpiperazinyl, Piperidinyl, Pyrrolyl, Pyrrolidinyl, Pyridyl, -NR_aR_b, gegebenenfalls substituierten Acylamino, gegebenenfalls substituierten Alkoxyformyl und Carbamoyl ausgewählt ist;
R⁴, R⁵, R⁶ und R⁷ jeweils unabhängig aus Wasserstoff, einem Halogen, einem C₁-C₆-Alkoxy, Hydroxy, gegebenenfalls substituierten Acyloxy, Amino, gegebenenfalls substituierten Acylamino, gegebenenfalls substituierten C₁-C₆-Alkyl, CN, einer Sulfonatgruppe, Sulfamoyl, Carbamoyl, Carboxy, gegebenenfalls substituierten Alkoxyformyl, gegebenenfalls substituierten Phenyl, gegebenenfalls substituierten N-Alkylpiperazinyl, gegebenenfalls substituierten Morpholinyl, gegebenenfalls substituierten Piperidinyl, gegebenenfalls substituierten Pyrrolyl, gegebenenfalls substituierten Pyrrolidinyl, -NR_aR_b, gegebenenfalls substituierten Pyridyl ausgewählt sind;
R_a und R_b aus einem Alkyl und einem Alkenyl ausgewählt sind; und
m eine ganze Zahl von 0 bis 3 ist.In a preferred embodiment of the present invention, the compound has the structure of general formula II:

in which
Y is selected from N, CH;
Z is selected from N, CR ⁶ ;
R ^{1 is} hydrogen, halogen, C ₁ -C ₆ alkoxy, an optionally substituted C ₁ -C ₆ alkyl, an optionally substituted aryl or an optionally substituted aralkyl;
R ^{3 is} independently selected from hydrogen, amino, hydroxy, optionally substituted acyloxy, alkoxy, halogen, optionally substituted alkyl, CN, a sulfonate group, sulfamoyl, carboxy, morpholinyl, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b optionally substituted acylamino, optionally substituted alkoxyformyl and carbamoyl;
R ⁴ , R ⁵ , R ⁶ and R ^{7 are} each independently selected from hydrogen, halo, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, a sulfonate group, sulfamoyl, carbamoyl, carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl are selected;
R _a and R _{b are selected} from an alkyl and an alkenyl; and
m is an integer from 0 to 3.

In a preferred embodiment of formula II, R ^{3 is selected} from an optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₄ alkyl, CN, a sulfonate group, sulfamoyl, carboxy and optionally substituted alkoxyformyl.

wobei
R¹ Wasserstoff, Halogen, C₁-C₆-Alkoxy, ein gegebenenfalls substituiertes C₁-C₆-Alkyl, ein gegebenenfalls substituiertes Aryl oder ein gegebenenfalls substituiertes Aralkyl ist;
R³ unabhängig aus Wasserstoff, Amino, Hydroxy, gegebenenfalls substituierten Acyloxy, Alkoxy, Halogen, gegebenenfalls substituierten Alkyl, CN, einer Sulfonatgruppe, Sulfamoyl, Carboxy, Morpholinyl, N-Alkylpiperazinyl, Piperidinyl, Pyrrolyl, Pyrrolidinyl, Pyridyl, -NR_aR_b, gegebenenfalls substituierten Acylamino, gegebenenfalls substituierten Alkoxyformyl und Carbamoyl ausgewählt ist;
R⁵, R⁶ und R⁷ jeweils unabhängig aus Wasserstoff, einem Halogen, einem C₁-C₆-Alkoxy, Hydroxy, gegebenenfalls substituierten Acyloxy, Amino, gegebenenfalls substituierten Acylamino, gegebenenfalls substituierten C₁-C₆-Alkyl, CN, einer Sulfonatgruppe, Sulfamoyl, Carbamoyl, Carboxy, gegebenenfalls substituierten Alkoxyformyl, gegebenenfalls substituierten Phenyl, gegebenenfalls substituierten N-Alkylpiperazinyl, gegebenenfalls substituierten Morpholinyl, gegebenenfalls substituierten Piperidinyl, gegebenenfalls substituierten Pyrrolyl, gegebenenfalls substituierten Pyrrolidinyl, -NR_aR_b, gegebenenfalls substituierten Pyridyl ausgewählt sind;
R_a und R_b aus einem Alkyl und einem Alkenyl ausgewählt sind; und
m = 0, 1, 2 oder 3 ist.In a particularly preferred embodiment of the present invention, the compound has the structure of general formula III:

in which
R ^{1 is} hydrogen, halogen, C ₁ -C ₆ alkoxy, an optionally substituted C ₁ -C ₆ alkyl, an optionally substituted aryl or an optionally substituted aralkyl;
R ^{3 is} independently selected from hydrogen, amino, hydroxy, optionally substituted acyloxy, alkoxy, halogen, optionally substituted alkyl, CN, a sulfonate group, sulfamoyl, carboxy, morpholinyl, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b optionally substituted acylamino, optionally substituted alkoxyformyl and carbamoyl;
R ⁵ , R ⁶ and R ^{7 are} each independently selected from hydrogen, halo, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, a sulfonate group, sulfamoyl, carbamoyl, carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, NR _a R _b , optionally substituted pyridyl are selected;
R _a and R _{b are selected} from an alkyl and an alkenyl; and
m = 0, 1, 2 or 3.

In a preferred embodiment of the formula III, R ^{1 is selected} from H and an alkyl.

In a preferred embodiment of the formula III, R ^{3 is selected} from an optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₄ alkyl, CN, a sulfonate group, sulfamoyl, carboxy and optionally substituted alkoxyformyl.

In a preferred embodiment of formula III, R ⁵ and R ^{6 are} independently selected from H, alkoxy, morpholinyl, halo, N-alkylpiperazinyl, piperidinyl, pyrrolidinyl, -NR _a R _b , acylamino and carbamoyl (NH ₂ C (O) -) wherein R _a and R _b are selected from alkyl and alkenyl.

In a preferred embodiment of formula III, R ^{5 is selected} from H, alkoxy, morpholinyl, halo, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b , acylamino and carbamoyl (NH ₂ C (O) -) where R _a and R _b can be selected from alkyl and alkenyl.

In a preferred embodiment of formula III, R ^{5 is selected} from H, alkoxy, morpholinyl, halo, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b , acylamino and carbamoyl (NH ₂ C (O) -) where R _a and R _b can be selected from alkyl and alkenyl, and R ⁶ = H.

In a preferred embodiment of formula III, R ^{5 is selected} from a halogen, 4-N-methylpiperazinyl, N-morpholinyl, N-piperidinyl, N-pyrrolyl, N-pyrrolidinyl, N, N-diethylamino, an N, N-dimethylmethylamine group and 4-pyridyl selected.

In a preferred embodiment of formula III, R ⁵ and R ^{6 are} H, and R ⁷ is acylamino.

In a preferred embodiment of the formula III, R ^{3 is} independently selected from acylamino, acyloxy and alkoxy.

In a preferred embodiment of the formula III, R ^{3 is} independently selected from the following groups:

wobei X ein Halogen ist.In a preferred embodiment of the formula III, R ^{3 is selected} from the following groups:

where X is a halogen.

In a preferred embodiment of the formula III, R ^{3 is selected} from the following groups:

In a preferred embodiment of the formula III, m = 1.

In a preferred embodiment of the formula III m = 1, and R ³ is located at position 4 of the phenyl group.

The present invention also encompasses the use of the compound of the present invention in the manufacture of drugs for the treatment of Epidermal Growth Factor Receptor Kinase (EGFR) mediated diseases.

The present invention also encompasses the use of the compound of the present invention in the manufacture of drugs for the treatment of B lymphocyte kinase (BLK) or FMS type tyrosine kinase 3 (FLT3) mediated diseases.

In one embodiment, the disease is cancer.

In one embodiment, the cancer is selected from diffuse B-cell lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, follicular lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, marginal zone lymphoma of the spleen, plasma cell myeloma, plasma cell tumors, extranodal marginal zone B. Cell lymphoma, marginal zone B-cell lymphoma of the lymph nodes, mantle cell lymphoma, thymic large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma, pain by mycosis fungoides, lymphoblastic lymphoma, T cell prolymphocytic leukemia, T cell granular lymphocytic leukemia, aggressive NK cell leukemia, cutaneous T cell lymphoma, plastic large cell lymphoma, peripheral T cell lymphoma, adult T cell lymphoma, acute myeloid leukemia , acute lymphocytic leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic myel leukemia, chronic neutrophilic leukemia, acute undifferentiated leukemia, degenerative developmental large cell lymphoma, prolymphocytic leukemia, juvenile myelomonocytic leukemia, adult T-cell ALL-AML mixed myelodysplasia in three cell lines, MLL (mixed lineage leukemia), myelodysplastic Syndrome, myeloproliferative disorders and multiple myeloma.

In one embodiment, the disease is an immune disorder.

In one embodiment, the immune disorder is selected from arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, ord thyroiditis, Graves' disease, Felty syndrome, multiple sclerosis , infectious neuronal inflammation, acute disseminated encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis vulgaris, graft-versus-host reaction, transplantation, allergic reaction to transfusion, allergy, type I hypersensitivity, allergic conjunctivitis, allergic rhinitis and atopic dermatitis.

In one embodiment, the cancer is from non-small cell lung cancer, breast cancer, prostate cancer, glial cell tumors, ovarian cancer, head and neck squamous cell carcinoma, cervix cancer, esophageal cancer, liver cancer, kidney cancer, pancreatic cancer, colon cancer, skin cancer, leukemia, lymphoma, gastric cancer, multiple myeloma and solid Tumors selected.

The present invention also encompasses the use of the compound of the present invention in the manufacture of epidermal growth factor receptor kinase (EGFR) inhibiting agents. The present invention also encompasses the use of the compound of the present invention in the production of B lymphocyte kinase (BLK) or FMS type tyrosine kinase 3 (FLT3) inhibitors.

In the present invention, the above uses of the compound of formula III are particularly preferred.

The present invention also includes a pharmaceutical composition comprising the compound according to the present invention, and the pharmaceutical composition may optionally comprise a pharmaceutically acceptable carrier, excipient, diluent and the like.

Practical implementation of the invention

The terms used here are defined as follows:
The term "alkyl" as used herein refers to a saturated straight or branched alkyl of 1 to 10 carbon atoms, and preferably alkyl includes an alkyl of 2 to 8 carbon atoms, 1 to 6 carbon atoms, 1 to 4 carbon atoms, 3 to 8 carbon atoms Carbon atoms, 1 to 3 carbon atoms. Examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, heptyl and the like.

Alkyl may be substituted with one or more (eg 2, 3, 4 or 5) substituents, for example with a halogen or haloalkyl. For example, the alkyl may be an alkyl substituted with 1 to 4 fluorine atoms or a fluorinated alkyl substituted alkyl.

The term "alkoxy" as used herein refers to an alkyl substituted oxy. A preferred alkoxy is an alkoxy having a length of 1 to 6 carbon atoms, more preferably an alkoxy having a length of 1 to 4 carbon atoms. Examples of alkoxy include methoxy, ethoxy, propoxy and the like.

As used herein, the term "alkenyl" generally means a monovalent hydrocarbon group having at least one double bond, generally comprising 2 to 8 carbon atoms, preferably 2 to 6 carbon atoms, and may be straight chain or branched. Examples of alkenyl include ethenyl, propenyl, isopropenyl, butenyl, isobutenyl, hexenyl and the like.

As used herein, the term "alkynyl" generally means a monovalent hydrocarbon group having at least one triple bond, generally comprising 2 to 8 carbon atoms, preferably 2 to 6 carbon atoms, more preferably 2 to 4 carbon atoms, and may be straight chain or branched. Examples of alkynyl groups are ethynyl, propynyl, isopropynyl, butynyl, isobutinyl, hexynyl and the like.

The term "halogen atom" or "halogen" as used herein refers to fluorine, chlorine, bromine and iodine.

"Aryl" means a monocyclic, bicyclic or tricyclic aromatic group having 6 to 14 carbon atoms and includes phenyl, naphthyl, phenanthryl, anthryl, indenyl, fluorenyl, tetralin, indanyl and the like. Aryl may optionally be substituted with 1 to 5 (eg 1, 2, 3, 4 or 5) substituents selected from halogen, C _1-4 aldehyde, C _1-6 alkyl, cyano, nitro, Amino, hydroxy, hydroxymethyl, a halo-substituted alkyl (e.g., trifluoromethyl), halo-substituted alkoxy (e.g., trifluoromethoxy), carboxy, C _1-4 alkoxy, ethoxyformyl, N (CH ₃ ), and a C _1-4 . Acyl, a heterocyclyl or a heteroaryl and the like are selected.

The term "aralkyl" as used herein refers to an aryl-substituted alkyl, for example, a phenyl-substituted C ₁ -C ₆ alkyl. Examples of aralkyl include arylmethyl, arylethyl, etc., such as benzyl, phenethyl, and the like.

For example, aryl may be substituted with 1 to 3 substituents selected from a halogen, -OH, C _1-4 alkoxy, C _1-4 alkyl, -NO ₂ , -NH ₂ , -N (CH ₃ ) ₂ , a carboxy and ethoxy-formyl and the like are selected.

The term "five- or six-membered heterocycle" as used herein includes, inter alia, heterocyclic groups comprising 1 to 3 heteroatoms selected from O, S and N, including (among others) furyl, thienyl, pyrrolyl, pyrrolidinyl, pyrazolyl, imidazolyl , Triazolyl, oxazolyl, pyranyl, pyridyl, pyrimidinyl, pyrazinyl, piperidinyl, morpholinyl and the like.

The term "heteroaryl" as used herein means that the group comprises 5 to 14 ring atoms and 6, 10 or 14 electrons coexist in the ring system. And the ring atoms contained are carbon atoms and 1 to 3 heteroatoms optionally selected from O, N, S. Suitable heteroaryl includes piperazinyl, morpholinyl, piperidinyl, pyrrolidinyl, thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, including but not limited to 2-pyridyl, 3-pyridyl and 4-pyridyl, pyrazinyl, pyrimidinyl and the like.

The heteroaryl or the five- or six-membered heterocycle may optionally be substituted with 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from a halogen, a C _1-4 aldehyde group, a straight chain or a straight chain branched C _1-6 alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, a halo-substituted alkyl (e.g., trifluoromethyl), a halo-substituted alkoxy (e.g., trifluoromethoxy), carboxy, C _1-4 alkoxy, ethoxyformyl , N (CH ₃ ) and C _1-4 acyl are selected.

The term "acyloxy" as used herein refers to a group having the structure of the formula "-O-C (O) -R" wherein R may be selected from an alkyl, alkenyl and alkynyl. And R may be optionally substituted.

wobei R_a und R_b aus einem Alkyl und einem Alkenyl ausgewählt sind, ausgewählt sein kann.As used herein, the term "acylamino" refers to a group having the structure of the formula "-R'-NH-C (O) -R" where R 'may be selected from a bond or an alkyl and R is an alkyl, Alkenyl, alkynyl, an NR _a R _b -substituted alkyl, an NR _a R _b -substituted alkenyl, an NR _a R _b -substituted alkynyl, a halo-substituted alkyl, a cyano-substituted alkenyl,

wherein R _a and R _b are selected from an alkyl and an alkenyl may be selected.

As used herein, "optionally substituted" means that the group modified by the term may optionally be substituted with 1 to 5 (eg, 1, 2, 3, 4 or 5) substituents selected from a halogen, a C _{1 4-} aldehyde group, a straight chain or branched C _1-6 alkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, a halo-substituted alkyl (eg trifluoromethyl), a halogen-substituted alkoxy (eg trifluoromethoxy), carboxy, C _1-4 alkoxy, ethoxyformyl, N (CH ₃ ) and C _1-4 acyl are selected.

According to the present invention there is provided a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula I, II or III according to the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

Examples of a pharmaceutically acceptable salt of the compound according to the present invention include a salt of an inorganic or organic acid such as a hydrochloride, hydrobromide, sulfate, citrate, lactate, tartrate, maleate, fumarate, mandelate and oxalate, and an inorganic salt or organic base such as sodium hydroxide, tris (hydroxymethyl) aminomethane (IRIS, amine tromethamine) and N-methylglucamine.

Each person will have different requirements, but the optimal dosage for each active ingredient in the pharmaceutical composition of the present invention may be determined by those skilled in the art. In general, the compound according to the present invention or a pharmaceutically acceptable salt thereof is orally administered to a mammal at a dose of about 0.0025 to 50 mg / kg of body weight per day, preferably about 0.01 to 10 mg / kg of body weight. For example, the oral dosage unit may comprise from about 0.01 to 50 mg, preferably from about 0.1 to 10 mg, of the compound of the invention. Dose units may be administered once or several times, one or more tablets a day, each containing about 0.1 to 50 mg, suitably about 0.25 to 10 mg, of the compound of the present invention or a solvate thereof.

The pharmaceutical composition of the present invention may be formulated into forms suitable for various routes of administration, including but not limited to parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, for the treatment of cancer and other diseases. nasal or topical route of administration. The amount administered causes relief or elimination of one or more conditions. For the treatment of a particular disease, the effective amount is sufficient to alleviate or reduce the symptoms of disease in any way. Such doses may be administered as a single dose or they may be administered according to an effective treatment. The dose can cure the disease but is usually given to relieve the symptoms of the disease. In general, repeated administration is required to achieve the desired relief of the symptoms. The dosage is determined depending on the age, health and weight of the patient, concurrent treatment, frequency of treatment and desired therapeutic effects.

Pharmaceutical compositions of the present invention can be administered to any mammal as long as the therapeutic effects can be achieved. Of the mammals, man is the most important.

The compound or pharmaceutical composition of the present invention may be useful for the treatment or prevention of various diseases mediated by the epidermal growth factor receptor kinase (EGFR). The disease mediated by EGFR is a variety of cancers. The cancers include non-small cell lung, breast, prostate, glial, ovarian, squamous cell carcinoma, cervix, esophageal, liver, kidney, pancreatic, colon, skin, leukemia, lymphoma, gastric, multiple myeloma, and solid tumors ,

The compound or pharmaceutical composition of the present invention can be used for the treatment of various diseases mediated by B-lymphocyte kinase (BLK) or FMS-like tyrosine kinase 3 (FLT3). The diseases mediated by BLK or FLT3 are various cancers and immune diseases. The cancer includes, but is not limited to, diffuse B-cell lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, follicular lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, spleen marginal zone lymphoma, plasma cell myeloma, plasma cell tumors, extranodal marginal zone B cell Lymphoma, marginal zone B-cell lymphoma of the lymph nodes, mantle cell lymphoma, thymic large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma, pain by mycosis fungoides, lymphoblastic lymphoma, T-cell -prolymphocytic leukemia, T-cell granular lymphocytic leukemia, aggressive NK cell leukemia, cutaneous T-cell lymphoma, plastic large cell lymphoma, peripheral T-cell lymphoma, adult T-cell lymphoma, acute myeloid leukemia, acute lymphocytic Leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic neutrophils n-leukemia, acute undifferentiated leukemia, degenerative developmental large cell lymphoma, prolymphocytic leukemia, juvenile myelomonocytic leukemia, adult T-cell ALL-AML mixed myelodysplasia in three cell lines, mixed lineage leukemia (MLL), myelodysplastic syndrome, myeloproliferative disorders and multiple myeloma. The immune diseases include arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, ord thyroiditis, Graves' disease, Felty's syndrome, multiple sclerosis, infectious neuronal Inflammation, acute disseminated encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis vulgaris, graft-versus-host reaction, transplantation, allergic reaction to transfusion, allergy, type I hypersensitivity, allergic conjunctivitis, allergic rhinitis and Atopic dermatitis.

The pharmaceutical preparations of the present invention can be prepared by a known method. For example, they can be prepared by a conventional mixing, granulating, drageeing, dissolving or freeze-drying method. During the preparation of oral preparations, solid excipients and active compounds may be combined and optionally ground. If necessary, an appropriate amount of excipient may be added and the mixture of granules may be made into tablets or dragee cores.

Suitable excipients (in particular fillers) are, for example, sugars, such as lactose or sucrose, mannitol or sorbitol; Cellulose preparations or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate; and binders, such as starch, including corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose or polyvinylpyrrolidone. If desired, disintegrants may be added, for example, the above-mentioned starches, as well as carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate. Auxiliaries, in particular flow improvers and lubricants, for. B. Silica, talc, stearic acid salts such as magnesium and calcium stearate, stearic acid or polyethylene glycol can be added. If necessary, suitable coatings that are resistant to gastric juice may be applied to the dragee core. For this purpose, concentrated sugar solutions can be applied. Such a solution may contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. For the preparation of enteric coatings, a suitable cellulose solution, for example, cellulose acetate phthalate or hydroxypropylmethylcellulose phthalate, may be used. Dyes or pigments may be added to the coating of tablets or dragee cores, for example to identify or characterize the dosage combinations of active agents.

Accordingly, the present invention also provides a method of treating or preventing EGFR-mediated diseases which comprises administering the compound or pharmaceutical composition of the present invention to a patient in need thereof.

The methods of administration include, but are not limited to, various methods of administration known in the art and which may be determined according to the actual situation of the patients. These methods include, but are not limited to, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, nasal, or topical routes.

The present invention also encompasses the use of the compound of the present invention in the manufacture of medicaments for the treatment of an EGFR, BLK or FLT3 mediated disease.

The present invention also encompasses the use of the compound of the present invention in the preparation of medicaments for the inhibition of kinases and methods for the inhibition of kinases. The method comprises administering an inhibitory amount or a therapeutically / prophylactically effective amount of a compound or pharmaceutical composition of the present invention to a patient in need thereof.

In the present invention, the patient may be a mammal and is preferably a human.

In the present invention, the kinase comprises, among others, EGFR, BLK, FLT3, HER2, HER4, FLT1, CDK2, JAK2, LCK, LYNA, cKit, PIM1, FGFR3, FGFR1, PDGFRa, PDGFRb, KDR, SRC, ABL, AURB, C-MET, BRAF, PKACa, IKKb, IGF1R, GSK3b, P38a and ERK1.

The present invention also encompasses the use of the compound of the present invention in the manufacture of medicaments for the inhibition of various diseases mediated by the kinases and methods for the treatment or prevention of various diseases mediated by the kinases. The methods include administering a therapeutically / prophylactically effective amount of the compound or pharmaceutical composition of the present invention to a patient in need thereof. Among these diseases mediated by kinase are, among others, the various cancers and immune diseases described above.

Synthesis of inhibitors

The present invention will be further illustrated by the following examples. These examples are intended to illustrate the present invention, but do not limit the scope of the present invention in any way.

Reagents and conditions: (a) ArNH ₂ , DIPEA, 1,4-dioxane, at room temperature; (b) ArNH ₂ , DIPEA, 1,4-dioxane, at room temperature; (c) Pd / C, H ₂ , EtOH; (d) R ² COCOOEt, HOAc, EtOH, at reflux; (e) trifluoroacetic acid, CH ₂ Cl ₂ , 0 ° C to RT; (f) acid chloride, Et ₃ N, CH ₂ Cl ₂ , 0 ° C to RT; or acid chloride, 1-methyl-2-pyrrolidone, CH ₃ CN, 0 ° C to RT.

In the above production process, R ¹ to R ⁴ are defined as described above. Various starting compounds routinely obtained as a raw material in the art may be used by those skilled in the art in accordance with actual needs to prepare the compound of the present invention.

example 1

The particular method for steps a to f, as mentioned above, is shown below:

Synthesis of tert-butyl (4- (2-chloro-5-nitropyrimidyl-4-amino) phenyl) carbamate (step a)

2,4-Dichloro-5-nitropyrimidine (95 mg, 0.49 mmol) was placed in a 10 mL round bottom flask, 3 mL of 1,4-dioxane was added and stirred at room temperature. Tert-butyl (4-aminophenyl) carbamate (100 mg, 0.48 mmol) and N, N-diisopropylethylamine (69 mg, 0.53 mmol) were dissolved in 2 mL of 1,4-dioxane. The resulting solution was added dropwise to the above reaction solution. After completion of the addition, the resulting mixture was stirred for 0.5 h at room temperature and TLC showed that the starting material was completely reacted. The solvent was removed by rotary evaporation and the crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10: 1, v / v) to give tert-butyl 4- (2-chloro-5-nitropyrimidyl-4 -amino) -phenyl) carbamate in the form of orange solids (144 mg, 82% yield).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.38 (s, 1H), 9.46 (s, 1H), 9.12 (s, 1H), 7.49 (d, J = 8.6 Hz, 2H), 7.39 (d, J = 8.6 Hz, 2H), 1.49 (s, 9H).

Synthesis of tert-butyl (4- (2- (4-methoxyphenylamino) -5-nitropyrimidyl-4-amino) phenyl) carbamate (step b)

Tert-Butyl (4- (2-chloro-5-nitropyrimidyl-4-amino) phenyl) carbamate (50 mg, 0.14 mmol), p-anisidine (17 mg, 0.14 mmol), N, N-diisopropylethylamine (18 mg, 0.18 mmol) were placed in a 10 mL round bottom flask, 5 mL of 1,4-dioxane was added, and stirred for 4 h at room temperature. TLC showed that the starting material was completely reacted. The solvent was removed by rotary evaporation and the crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 4: 1, v / v) to give tert -butyl (4- (2- (4-methoxyphenylamino) -5- nitropyrimidyl-4-amino) phenyl) carbamate as a yellow solid (51 mg, yield 82%).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.30 (s, 1H), 10.26 (s, 1H), 9.45 (s, 1H), 9.04 (s, 1H) , 7.49 (d, J = 8.8 Hz, 2H), 7.45 (d, J = 8.8 Hz, 2H), 7.40 (d, J = 8.6 Hz, 2H), 6 , 75 (d, J = 8.6 Hz, 2H), 3.73 (s, 3H), 1.50 (s, 9H).

Synthesis of tert-butyl (4- (5-amino-2- (4-methoxyphenylamino) pyrimidyl-4-amino) phenyl) carbamate (step c)

Tert-butyl (4- (2- (4-methoxyphenylamino) -5-nitropyrimidyl-4-amino) phenyl) carbamate (45mg, 0.10mmol) was placed in a 50ml round bottom flask. 20 ml of ethanol and 5 mg of palladium on carbon (10% Pd) were added, hydrogen was charged, and the resulting reaction system was stirred at room temperature overnight. After completion of the reaction, the system was filtered and the filtrate was spun dry. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 5: 1, v / v) to give tert -butyl (4- (5-amino-2- (4-methoxyphenylamino) pyrimidyl-4-amino) phenyl) carbamate as a pale pink solid (30 mg, yield 83%).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 9.23 (s, 1H), 8.42 (s, 1H), 8.10 (s, 1H), 7.62 (d, J = 9.2 Hz, 2H), 7.56 (s, 1H), 7.53 (d, J = 9.2 Hz, 2H), 7.40 (d, J = 8.8 Hz, 2H), 6 , 77 (d, J = 8.8 Hz, 2H), 3.70 (s, 3H), 1.48 (s, 9H).

Synthesis of tert-butyl (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) carbamate (step d)

Tert-butyl (4- (5-amino-2- (4-methoxyphenylamino) pyrimidinyl-4-amino) phenyl) carbamate (30 mg, 0.07 mmol) was added to a 10 mL round bottom flask. To this was added 0.29 ml of glacial acetic acid and 5 ml of anhydrous ethanol and then ethyl glyoxylate (50% in toluene) (16 mg, 0.08 mmol) was added. The resulting reaction mixture was heated to reflux and stirred overnight. Upon completion, solids precipitated. The solid was filtered off and the filter cake was washed with ethanol, ammonia water and deionized water and then dried to give tert -butyl (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl). carbamate as a yellow solid gave (18 mg, 76% yield).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.08 (s, 1H), 9.64 (s, 1H), 8.84 (s, 1H), 8.03 (s, 1H) , 7.65 (d, J = 8.4 Hz, 2H), 7.30-7.28 (m, 4H), 6.61 (br, 2H), 3.67 (s, 3H), 1, 52 (s, 9H).

Synthesis of 8- (4-aminophenyl) -2- (4-methoxyphenyl) -7 (8H) -pteridinone (Compound 001) (Step e)

Tert-butyl (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) carbamate (18mg, 0.04mmol) was added to a 5ml round bottom flask. 2 ml of dichloromethane was added and the resulting mixture was stirred at 0 ° C. 0.5 ml of trifluoroacetic acid was added and the resulting mixture was stirred at 0 ° C for 1 hour and then at room temperature for an additional 1 hour. After the reaction was completed, a saturated sodium hydrogencarbonate solution was added to make the solution alkaline, and the resulting mixture was extracted with dichloromethane (3 × 50 ml). The organic layer was washed with deionized water and a saturated sodium chloride solution and then dried over anhydrous sodium sulfate. The solvent was removed by spin-drying to give 8- (4-aminophenyl) -2- (4-methoxyphenyl) -7 (8H) -pteridinone as a yellow solid (14 mg, 99% yield).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.04 (br, 1H), 8.81 (s, 1H), 8.00 (s, 1H), 7.40 (d, J = 7.6Hz, 2H), 6.98 (d, J = 8.4Hz, 2H), 6.73 (d, J = 8.4Hz, 2H), 6.67 (br, 2H), 5 , 44 (s, 2H), 3.70 (s, 3H). ¹³ C-NMR (100 MHz, DMSO-d ₆ ): δ 159.19, 158.53, 157.17, 154.95, 151.76, 149.66, 146.68, 133.17, 129.22 , 122.66, 121.04, 120.70, 114.37, 113.87, 55.55. HRMS (ESI) calculated for C ₁₉ H ₁₇ N ₆ O ₂ [M + H] ⁺ 361.1413, found 361.1414.

Synthesis of N- (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) -acrylamide (Compound 002) (Step f)

8- (4-Aminophenyl) -2- (4-methoxyphenyl) -7 (8H) -pteridinone (100 mg, 0.28 mmol) was placed in a 100 mL round bottom flask. 50 ml of dichloromethane and triethylamine (28 mg, 0.28 mmol) were added and the resulting mixture was stirred at 0 ° C. Acryloyl chloride (29 mg, 0.31 mmol) was dissolved in 5 ml of dichloromethane and added dropwise to the above reaction solution. After the addition, the resulting reaction mixture was stirred at room temperature overnight. The solvent was removed by rotary evaporation and the crude product was purified by silica gel column chromatography (dichloromethane / ethyl acetate = 5: 1, v / v) to give N- (4- (2- (4-methoxyphenylamino) -7-oxo -8 (7H) -pteridinyl) phenyl) acrylamide as a yellow solid (34 mg, yield 30%).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.42 (s, 1H), 10.07 (br, 1H), 8.84 (s, 1H), 8.04 (s, 1H) , 7.87 (d, J = 8.8 Hz, 2H), 7.38 (d, J = 8.8 Hz, 2H), 7.30 (br, 2H), 6.59 (br, 2H) , 6.52 (dd, J = 17.0, 10.0 Hz, 1H), 6.33 (dd, J = 17.0, 1.8 Hz, 1H), 5.82 (dd, J = 10 , 0, 1.8 Hz, 1H), 3.62 (s, 3H). ¹³ C NMR (100 MHz, DMSO-d ₆ ): δ 163.90, 159.28, 158.51, 156.68, 155.02, 151.44, 146.65, 139.72, 133.00, 130.18, 129.50, 127.72, 121.02, 120.61, 113.77, 55.40. HRMS (ESI) calculated for C ₂₂ H ₁₉ N ₆ O ₃ [M + H] ⁺ 415.1519, found 415.1515.

¹H-NMR (400 MHz, DMSO-d₆): δ 10,44 (s, 1H), 10,00 (s, 1H), 8,82 (s, 1H), 8,02 (s, 11-1), 7,88 (d, J = 8,0 Hz, 1H), 7,36 (d, J = 8,4 Hz, 1H), 7,22 (br, 2H), 6,59 (br, 2H), 6,52 (dd, J = 17,2, 10,2 Hz, 1H), 6,33 (d, J = 17,2 Hz, 1H), 5,85 (d, J = 10,2 Hz, 1H), 3,67 (br, 4H), 2,92 (br, 4H). HRMS (ESI) berechnet für C₂₅H₂₄N₇O₃ [M+H]⁺ 470,1941, gefunden 470,1932. N-(4-(2-(4-Methoxyphenylamino)-6-methyl-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 004)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,44 (s, 1H), 9,90 (br, 1H), 8,77 (s, 1H), 7,87 (d, J = 8,8 Hz, 2H), 7,50 (d, J = 8,8 Hz, 2H), 7,29 (br, 2H), 6,59 (br, 2H), 6,52 (dd, J 17,0, 10,0 Hz, 1H), 6,33 (dd, J = 17,0, 1,9 Hz, 1H), 5,82 (dd, J = 10,0, 1,9 Hz, 1H), 3,61 (s, 3H), 2,42 (s, 3H). HRMS (ESI) berechnet für C₂₃H₂₁N₆O₃ [M+H]⁺ 429,1675, gefunden 429,1671. 8-(3-Aminophenyl)-2-(4-methoxyphenyl)-7(8H)-pteridinon (Verbindung 005)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,06 (br, 1H), 8,83 (s, 1H), 8,01 (s, 1H), 7,41 (d, J = 8,0 Hz, 214), 7,22 (t, J = 8,0 Hz, 1H), 6,75 (d, J = 7,6 Hz, 1H), 6,67 (br, 2H), 6,53 (s, 1H), 6,48 (d, J = 7,6 Hz, 1H), 5,35 (s, 2H), 3,69 (s, 3H). HRMS (ESI) berechnet für C₁₉H₁₇N₆O₂ [M+H]⁺ 361,1413, gefunden 361,1413. N-(3-(2-(4-Methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 006)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,42 (s, 1H), 10,10 (br, 1H), 8,85 (s, 1H), 8,05 (s, 1H), 7,84 (d, J = 8,0 Hz, 1H), 7,78 (s, 1H), 7,56 (t, J = 8,0 Hz, 1H), 7,31 (br, 2H), 7,13 (d, J = 8,0 Hz, 1H), 6,58 (br, 2H), 6,45 (dd, J = 16,8, 10,4 Hz, 1H), 6,26 (dd, J = 16,8, 1,6 Hz, 1H), 5,77 (dd, J = 10,4, 1,6 Hz, 1H), 3,65 (s, 3H). HRMS (ESI) berechnet für C₂₂H₁₉N₆O₃ [M+H]⁺ 415,1519, gefunden 415,1516. N-(3-(2-(4-Methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)propionamid (Verbindung 007)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,13 (s, 1H), 10,09 (s, 1H), 8,85 (s, 1H), 8,04 (s, 1H), 7,74 (d, J = 8,0 Hz, 1H), 7,71 (s, 1H), 7,53 (t, J = 8,0 Hz, 1H), 7,31 (br, 2H), 7,07 (d, J = 8,0 Hz, 1H), 6,59 (br, 2H), 3,67 (s, 3H), 2,33 (q, J = 7,6 Hz, 2H), 1,07 (t, J = 7,6 Hz, 311). HRMS (ESI) berechnet für C₂₂H₂₁N₆O₃ [M+H]⁺ 417,1675, gefunden 417,1678. N-(4-(2-(4-Methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)propionamid (Verbindung 008)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,15 (s, 1H), 10,08 (br, 1H), 8,85 (s, 1H), 8,04 (s, 1H), 7,80 (d, J = 8,4 Hz, 2H), 7,35–7,33 (m, 4H), 6,61 (br, 2H), 3,67 (s, 3H), 2,41 (q, J = 7,6 Hz, 2H), 1,14 (t, J = 7,6 Hz, 3H). HRMS (ESI) berechnet für C₂₂H₂₁N₆O₃ [M+H]⁺ 417,1675, gefunden 417,1674. 4-(Dimethylamino)-N-(4-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)-phenyl)-2-butenamid (Verbindung 009)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,45 (s, 1H), 10,10 (br, 1H), 8,85 (s, 1H), 8,05 (s, 1H), 7,87 (d, J = 8,8 Hz, 2H), 7,37 (d, J = 8,8 Hz, 2H), 7,30 (br, 2H), 6,82 (td, J = 15,4, 6,0 Hz, 1H), 6,60 (br, 2H), 6,40 (d, J = 15,4 Hz, 1H), 3,63 (s, 3H), 3,27 (d, J = 5,2 Hz, 2H), 2,33 (s, 6H). HRMS (ESI) berechnet für C₂₅H₂₆N₇O₃ [M+H]⁺ 472,2097, gefunden 472,2095. 4-(Dimethylamino)-N-(3-(2-(4-methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)-phenyl)-2-butenamid (Verbindung 010)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,33 (s, 1H), 10,08 (br, 1H), 8,86 (s, 1H), 8,05 (s, 1H), 7,83 (d, J 8,0 Hz, 1H), 7,78 (s, 1H), 7,55 (t, J = 8,0 Hz, 1H), 7,32 (br, 2H), 7,11 (d, J = 8,0 Hz, 1H), 6,74 (td, J 15,2, 5,6 Hz, 1H), 6,59 (br, 2H) 6,30 (d, J = 15,2 Hz, 1H), 3,66 (s, 3H), 3,06 (d, J 5,6 Hz, 2H), 2,17 (s, 6H). HRMS (ESI) berechnet für C₂₅H₂₄N₇O₃ [M+H]⁺ 472,2097, gefunden 472,2094. 4-(2-(4-Methoxyphenylamino)-7-oxo-8(7H)-pteridinyl)phenylacrylat (Verbindung 011)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,15 (s, 1H), 8,87 (s, 1H), 8,06 (s, 11-1), 7,51 (d, J = 8,8 Hz, 2H), 7,45 (d, J = 8,8 Hz, 2H), 7,31 (br, 2H), 6,69 (br, 2H), 6,60 (dd, J = 17,2, 1,6 Hz, 1H), 6,51 (dd, J = 17,2, 9,9 Hz, 1H), 6,22 (dd, J = 9,9, 1,6 Hz, 1H), 3,67 (s, 3H). HRMS (ESI) berechnet für C₂₂H₁₈N₅O₄ [M+H]⁺ 416,1359, gefunden 416,1359. 4-(Dimethylamino)-N-(4-(7-oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)-2-butenamid (Verbindung 012)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,37 (s, 11-1), 10,19 (br, 1H), 8,90 (s, 1H), 8,08 (s, 1H), 7,87 (d, J 8,4 Hz, 2H), 7,42 (d, J = 7,6 Hz, 2H), 7,38 (d, J = 8,4 Hz, 2H), 7,03 (br, 1H), 6,88 (t, J = 7,6 Hz, 1H), 6,82 (td, J = 15,4, 5,6 Hz, 1H), 6,37 (d, J = 15,4 Hz, 1H), 3,14 (d, J = 5,6 Hz, 2H), 2,24 (s, 6H). HRMS (ESI) berechnet für C₂₄H₂₄N₇O2 [M+H]⁺ 442,1991, gefunden 442,1989. 4-(Dimethylamino)-N-(3-(7-oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)-2-butenamid (Verbindung 013)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,32 (s, 1H), 10,17 (s, 1H), 8,90 (s, 1H), 8,08 (s, 1H), 7,81–7,79 (m, 2H), 7,55 (t, J = 8,0 Hz, 1H), 7,41 (d, J = 7,2 Hz, 2H), 7,12 (d, J = 8,0 Hz, 1H), 7,01 (br, 21-1), 6,87 (t, J = 7,2 Hz, 1H), 6,73 (td, J = 15,2, 5,6 Hz, 1H), 6,28 (d, J = 15,2 Hz, 1H), 3,05 (d, J = 5,6 Hz, 2H), 2,16 (s, 6H). HRMS (ESI) berechnet für C₂₄H₂₄N₇O₂ [M+H]⁺ 442,1991, gefunden 442,1996. N-(4-(7-Oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 014)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,44 (s, 1H), 10,19 (br, 1H), 8,90 (s, 1H), 8,09 (s, 1H), 7,88 (d, J = 8,4 Hz, 2H), 7,41–7,38 (m, 4H), 7,03 (br, 2H), 6,88 (t, J = 7,2 Hz, 1H), 6,53 (dd, J = 16,8, 10,4 Hz, 1H), 6,35 (dd, J = 16,8, 1,6 Hz, 1H), 5,84 (dd, J = 10,4, 1,6 Hz, 1H). HRMS (ESI) berechnet für C₂₁H₁₇N₆O₂ [M+H]⁺ 385,1413, gefunden 385,1405. N-(3-(7-Oxo-2-(phenylamino)-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 015)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,42 (s, 1H), 10,19 (s, 1H), 8,91 (s, 1H), 8,09 (s, 1H), 7,84–7,81 (m, 2H), 7,57 (t, J = 8,0 Hz, 1H), 7,41 (br, 2H), 7,15 (d, J = 7,6 Hz, 1H), 7,02 (br, 2H), 6,87 (t, J = 7,6 Hz, 1H), 6,45 (dd, J = 16,8, 10,4 Hz, 1H), 6,26 (dd, J = 16,8, 1,6 Hz, 1H), 5,77 (dd, J = 10,4, 1,6 Hz, 1H). HRMS (ESI) berechnet für C₂₁H₁₇N₆O₂ [M+H]⁺ 385,1413, gefunden 385,1413. N-(4-(2-(4-Chlorphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 016)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,46 (s, 1H), 10,34 (s, 1H), 8,92 (s, 1H), 8,11 (5, 1H), 7,88 (d, J = 8,8 Hz, 2H), 7,41–7,36 (m, 4H), 7,06 (br, 2H), 6,53 (dd, J = 16,8, 10,4 Hz, 1H), 6,36 (dd, J = 16,8, 1,6 Hz, 1H), 5,84 (dd, J = 10,4, 1,6 Hz, 1H). HRMS (ESI) berechnet für C₂₁H₁₆N₆O₂Cl [M+H]⁺ 419,1023, gefunden 419,1031. N-(3-(2-(4-Chlorphenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 017)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,44 (s, 1H), 10,34 (br, 1H), 8,93 (s, 1H), 8,11 (s, III), 7,84 (s, III), 7,81 (d, J = 8,4 Hz, 11-I), 7,59 (t, J = 8,0 Hz, 1H), 7,43 (d, J = 7,2 Hz, 2H), 7,15 (d, J 7,6 Hz, 1H), 6,46 (dd, J = 16,8, 10,4 Hz, 1H), 6,26 (dd, J = 16,8, 1,8 Hz, 1H), 5,77 (dd, J 10,12, 1,8 Hz, 1H). HRMS (ESI) berechnet für C₂₁H₁₆N₆O₂Cl [M+H]⁺ 419,1023, gefunden 419,1027. N-(3-(2-(4-Morpholinophenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 018)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,43 (s, 1H), 10,06 (s, 1H), 8,84 (s, 1H), 8,03 (s, 1H), 7,92 (br, 1H), 7,72 (s, 1H), 7,56 (t, J = 7,6 Hz, 1H), 7,27 (br, 2H), 7,12 (d, J = 7,2 Hz, 1H), 6,58 (br, 2H), 6,45 (dd, J = 16,8, 10,4 Hz, 1H), 6,26 (d, J = 16,8 Hz, 1H), 5,78 (d, J = 10,4 Hz, 1H), 3,71 (br, 4H), 2,94 (br, 4H). HRMS (ESI) berechnet für C₂₅H₂₄N₇O₃ [M+H]⁺ 470,1941, gefunden 470,1939. N-(4-(2-(4-(4-Methyl-1-piperazinyl)phenylamino)-7-oxo-8(7H)-pteridinyl)-phenyl)acrylamid (Verbindung 019)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,51 (s, 1H), 10,06 (s, 11-1), 8,83 (s, 1H), 8,03 (s, 1H), 7,89 (d, J = 8,4 Hz, 2H), 7,37 (d, J = 8,4 Hz, 2H), 7,17 (d, J = 6,4 Hz, 1H), 6,56–6,49 (m, 3H), 6,34 (d, J = 16,8 Hz, 1H), 5,85 (d, J = 10,8 Hz, 1H), 2,94 (br, 4H), 2,37 (br, 4H), 2,20 (s, 3H). HRMS (ESI) berechnet für C₂₆H₂₇N₈O₂ [M+H]⁺ 483,2257, gefunden 483,2259. N-(3-(2-(4-(4-Methyl-1-piperazinyl)-phenylamino)-7-oxo-8(7H)-pteridinyl)-phenyl)acrylamid (Verbindung 020)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,45 (s, 1H), 10,06 (s, 1H), 8,84 (s, 1H), 8,04 (s, 1H), 7,93 (br, 1H), 7,73 (s, 1H), 7,56 (t, J = 8,0 Hz, 1H), 7,25 (br, 2H), 7,12 (d, J = 8,0 Hz, 1H), 6,57 (br, 2H), 6,46 (dd, J = 16,8, 10,4 Hz, 111), 6,27 (dd, J = 16,8, 1,8 Hz, 1H), 5,78 (dd, J = 10,4, 1,8 Hz, 1H), 2,98 (br, 4H), 2,42 (br, 4H), 2,22 (s, 3H). HRMS (ESI) berechnet für C₂₆H₂₇N₈O₂ [M+H]⁺ 483,2257, gefunden 483,2259. N-(3-(7-Oxo-2-(4-(1-piperidinyl)phenylamino)-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 021)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,44 (s, 1H), 10,03 (s, 1H), 8,83 (s, 1H), 8,02 (s, 1H), 7,94 (br, 1H), 7,73 (s, 1H), 7,55 (t, J = 8,0 Hz, 111), 7,24 (br, 2H), 7,11 (d, J 8,0 Hz, 1H), 6,57 (br, 2H), 6,46 (dd, J = 17,0, 10,2 Hz, 1H), 6,26 (dd, J = 17,0, 1,8 Hz, 1H), 5,77 (dd, J = 10,2, 1,8 Hz, 1H), 2,95 (br, 4H), 1,57 (br, 4H), 1,49 (br, 2H). HRMS (ESI) berechnet für C₂₆H₂₆N₇O₂ [M+H]⁺ 468,2148, gefunden 468,2146. N-(3-(7-Oxo-2-(4-(1-pyrrolidinyl)phenylamino)-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 022)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,40 (s, 1H), 9,92 (s, 1H), 8,79 (s, 1H), 7,99 (s, HI), 7,90 (br, 1H), 7,74 (br, 1H), 7,54 (t, J = 8,0 Hz, 1H), 7,20 (br, 2H), 7,10 (d, J = 8,0 Hz, 1H), 6,46 (dd, J = 17,0, 10,2 Hz, 1H), 6,26 (dd, J = 17,0, 1,8 Hz, 1H), 6,20 (br, 2H), 5,77 (dd, J = 10,2, 1,8 Hz, 1H), 3,10 (br, 4H), 1,91 (br, 4H). HRMS (ESI) berechnet für C₂₅H₂₄N₇O₂ [M+H]⁺ 454,1991, gefunden 454,1995. N-(3-(2-(4-(Diethylamino)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 023)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,42 (s, 1H), 9,92 (s, 1H), 8,80 (s, 11-1), 8,00 (s, 1H), 7,92 (br, 1H), 7,73 (s, 1H), 7,53 (t, J = 8,0 Hz, 1H), 7,19 (br, 2H), 7,09 (d, J = 8,0 Hz, 1H), 6,46 (dd, J = 17,0, 10,2 Hz, 1H), 6,32 (br, 2H), 6,27 (dd, J = 17,0, 1,8 Hz, 1H), 5,76 (dd, J = 10,2, 1,8 Hz, 1H), 3,20 (br, 4H), 1,00 (t, J = 6,8 Hz, 6H). HRMS (ESI) berechnet für C₂₅H₂₆N₇O₂ [M+H]⁺ 456,2148, gefunden 456,2143. N-(3-(2-(4-(Acetylamino)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 024)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,43 (s, 1H), 10,16 (br, 1H), 9,78 (s, 1H), 8,87 (s, III), 8,06 (s, 114), 7,82 (d, J = 8,0 Hz, 1H), 7,79 (s, 1H), 7,56 (t, J = 8,0 Hz, 1H), 7,32 (br, 21-1), 7,23 (br, 21-1), 7,15 (d, J = 8,0 Hz, 1H), 6,46 (dd, J = 17,0, 10,2 Hz, 1H), 6,25 (dd, J = 17,0, 1,8 Hz, 1H), 5,76 (dd, J = 10,2, 1,8 Hz, 1H), 1,98 (s, 3H). HRMS (ESI) berechnet für C₂₃H₂₀N₇O₃ [M+H]⁺ 442,1628, gefunden 442,1624. 4-(8-(3-Acrylamidophenyl)-7-oxo-7,8-dihydropteridinyl-2-amino)benzamid (Verbindung 025)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,43 (s, 1H), 10,40 (s, 1H), 8,95 (s, 1H), 8,13 (s, 1H), 7,86 (s, 1H), 7,79 (d, J = 8,0 Hz, 1H), 7,71 (br, 1H), 7,61 (t, J 8,0 Hz, 1H), 7,55 (d, J = 7,6 Hz, 2H), 7,47 (br, 2H), 7,18 (d, J = 7,6 Hz, 2H), 6,44 (dd, J = 17,0, 10,2 Hz, 1H), 6,25 (dd, J = 17,0, 1,8 Hz, 1H), 5,76 (dd, J = 10,2, 1,8 Hz, 1H). HRMS (ESI) berechnet für C₂₂H₁₈N₇O₃ [M+H]⁺ 428,1471, gefunden 428,1476. N-(3-(2-(4-Methoxyphenylamino)-6-methyl-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 026)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,42 (s, 1H), 9,93 (br, 1H), 8,78 (s, 1H), 7,83 (d, J = 8,0 Hz, 1H), 7,77 (s, 11-1), 7,56 (t, J = 8,0 Hz, 1H), 7,31 (br, 2H), 7,11 (d, J = 8,0 Hz, 1H), 6,58 (br, 2H), 6,45 (dd, J = 17,0, 10,2 Hz, 1H), 6,26 (d, J r 17,0 Hz, 11-1), 5,77 (d, J = 10,2 Hz, 1H), 3,65 (s, 3H), 2,42 (s, 3H). HRMS (ESI) berechnet für C₂₃H₂₁N₆O₃ [M+H]⁺ 429,1675, gefunden 429,1675. N-(3-(8-(4-Methoxyphenyl)-7-oxo-7,8-dihydropteridin-2-amino)phenyl)acrylamid (Verbindung 027)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,17 (s, 1H), 10,01 (s, 1H), 8,89 (s, 1H), 8,07 (s, 1H), 7,63 (br, 1H), 7,33 (d, J = 8,8 Hz, 2H), 7,27 (d, J = 8,0 Hz, 1H), 7,23 (d, J = 8,0 Hz, 1H), 7,11 (d, J = 8,8 Hz, 2H), 6,89 (br, 1H), 6,46 (dd, J = 17,0, 10,2 Hz, 1H), 6,25 (dd, J = 17,0, 1,8 Hz, 1H), 5,74 (dd, J = 10,2, 1,8 Hz, 1H), 3,85 (s, 3H). HRMS (ESI) berechnet für C₂₂H₁₉N₆O₃ [M+H]⁺ 415,1519, gefunden 415,1519. 2-(3-Aminophenylamino)-8-(4-methoxyphenyl)-7(8H)-pteridinon (Verbindung 028)

¹H-NMR (400 MHz, DMSO-d₆): δ 9,91 (s, 1H), 8,85 (s, 1H), 8,04 (s, 1H), 7,35 (d, J 8,8 Hz, 2H), 7,16 (d, J = 8,8 Hz, 2H), 6,68–6,65 (m, 3H), 6,16 (d, J = 7,2 Hz, 1H), 4,63 (s, 2H), 3,86 (s, 3H). HRMS (ESI) berechnet für C₁₉H₁₇N₆O₂ [M+H]⁺ 361,1413, gefunden 361,1411. N-(4-(8-(4-Methoxyphenyl)-7-oxo-7,8-dihydropteridin-2-amino)phenyl)acrylamid (Verbindung 029)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,19 (br, 1H), 10,03 (s, 1H), 8,87 (s, 1H), 8,05 (s, 1H), 7,36–7,34 (m, 6H), 7,16 (d, J = 8,4 Hz, 2H), 6,41 (dd, J = 17,0, 10,2 Hz, 1H), 6,23 (dd, J = 17,0, 1,6 Hz, 1H), 5,72 (dd, J = 10,2, 1,6 Hz, 1H), 3,92 (s, 1H). HRMS (ESI) berechnet für C₂₂H₁₉N₆O₃ [M+H]⁺ 415,1519, gefunden 415,1524. 2-(4-Aminophenylamino)-8-(4-methoxyphenyl)-7(8H)-pteridinon (Verbindung 030)

¹H-NMR (400 MHz, DMSO-d₆): δ 9,87 (s, 1H), 8,77 (s, 1H), 7,97 (s, 1H), 7,32 (d, J = 8,8 Hz, 21-I), 7,13 (d, J = 8,8 Hz, 2H), 7,08 (br, 2H), 6,24 (br, 2H), 4,84 (s, 2H), 3,88 (s, 3H). HRMS (ESI) berechnet für C₁₉H₁₇N₆O₂ [M+H]+ 361,1413, gefunden 361,1417. N-(4-(2-(2-Methoxy)-4-(4-methoxy-1-piperazinyl)-phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 031)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,43 (s, 1H), 8,80 (s, 1H), 8,42 (s, 1H), 8,03 (s, 1H), 7,85 (d, J = 8,6 Hz, 2H), 7,34 (d, J = 8,6 Hz, 2H), 7,25 (d, J = 8,8 Hz, 1H), 6,54–6,48 (m, 21-1), 6,33 (dd, J = 17,0, 1,6 Hz, 1H), 6,02 (br, 1H), 5,84 (dd, J = 10,2, 1,6 Hz, 1H), 3,76 (s, 31-1), 3,02 (br, 4H), 2,43 (br, 4H), 2,23 (s, 3H). HRMS (ESI) berechnet für C₂₇H₂₉N₈O₃ [M+FI]⁺ 513,2363, gefunden 513,2362. N-(3-(2-(2-Methoxy-4-(4-methyl-1-piperazinyl)phenylamino)-7-oxo-8(7H)-pteridinyl)phenyl)acrylamid (Verbindung 032)

¹H-NMR (400 MHz, DMSO-d₆): δ 10,41 (s, 1H), 8,80 (s, 1H), 8.44 (br, 1H), 8,0 2 (s, 1H), 7,86 (br, 1H), 7,71 (s, 1H), 7,52 (t, J = 8,0 Hz, 1H), 7,30 (d, J = 7,6 Hz, 1H), 7,09 (d, J = 8,0 Hz, 1H), 6,53 (s, 1H), 6,46 (dd, J = 17,0, 10,2 Hz, 1H), 6,26 (dd, J = 17,0, 1,8 Hz, 1H), 6,02 (br, 1H), 5,78 (dd, J = 10,2, 1,8 Hz, 1H), 3,76 (s, 3H), 3,04 (br, 4H), 2,44 (br, 4H), 2,23 (s, 3H). HRMS (ESI) Berechnet für C₂₇H₂₉N₈O₃ [M+H]⁺ 513,2363, gefunden 513,2361.The following compounds were synthesized according to steps a-f above: N- (4- (2- (4-morpholinophenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) -acrylamide (Compound 003)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.44 (s, 1H), 10.00 (s, 1H), 8.82 (s, 1H), 8.02 (s, 11- 1), 7.88 (d, J = 8.0Hz, 1H), 7.36 (d, J = 8.4Hz, 1H), 7.22 (br, 2H), 6.59 (br, 2H), 6.52 (dd, J = 17.2, 10.2Hz, 1H), 6.33 (d, J = 17.2Hz, 1H), 5.85 (d, J = 10.2 Hz, 1H), 3.67 (br, 4H), 2.92 (br, 4H). HRMS (ESI) calculated for C ₂₅ H ₂₄ N ₇ O ₃ [M + H] ⁺ 470.1941, found 470.1932. N- (4- (2- (4-Methoxyphenylamino) -6-methyl-7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (Compound 004)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.44 (s, 1H), 9.90 (br, 1H), 8.77 (s, 1H), 7.87 (d, J = 8.8 Hz, 2H), 7.50 (d, J = 8.8 Hz, 2H), 7.29 (br, 2H), 6.59 (br, 2H), 6.52 (dd, J 17 , 0, 10.0 Hz, 1H), 6.33 (dd, J = 17.0, 1.9 Hz, 1H), 5.82 (dd, J = 10.0, 1.9 Hz, 1H) , 3.61 (s, 3H), 2.42 (s, 3H). HRMS (ESI) calculated for C ₂₃ H ₂₁ N ₆ O ₃ [M + H] ⁺ 429.1675, found 429.1671. 8- (3-aminophenyl) -2- (4-methoxyphenyl) -7 (8H) -pteridinone (Compound 005)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.06 (br, 1H), 8.83 (s, 1H), 8.01 (s, 1H), 7.41 (d, J = 8.0 Hz, 214), 7.22 (t, J = 8.0 Hz, 1H), 6.75 (d, J = 7.6 Hz, 1H), 6.67 (br, 2H), 6 , 53 (s, 1H), 6.48 (d, J = 7.6 Hz, 1H), 5.35 (s, 2H), 3.69 (s, 3H). HRMS (ESI) calculated for C ₁₉ H ₁₇ N ₆ O ₂ [M + H] ⁺ 361.1413, found 361.1413. N- (3- (2- (4-Methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (Compound 006)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.42 (s, 1H), 10.10 (br, 1H), 8.85 (s, 1H), 8.05 (s, 1H) , 7.84 (d, J = 8.0 Hz, 1H), 7.78 (s, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.31 (br, 2H) , 7.13 (d, J = 8.0 Hz, 1H), 6.58 (br, 2H), 6.45 (dd, J = 16.8, 10.4 Hz, 1H), 6.26 (dd, J = 16.8, 1.6 Hz, 1H), 5.77 (dd, J = 10.4, 1.6 Hz, 1H ), 3.65 (s, 3H). HRMS (ESI) calculated for C ₂₂ H ₁₉ N ₆ O ₃ [M + H] ⁺ 415.1519, found 415.1516. N- (3- (2- (4-Methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) propionamide (Compound 007)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.13 (s, 1H), 10.09 (s, 1H), 8.85 (s, 1H), 8.04 (s, 1H) , 7.74 (d, J = 8.0 Hz, 1H), 7.71 (s, 1H), 7.53 (t, J = 8.0 Hz, 1H), 7.31 (br, 2H) , 7.07 (d, J = 8.0 Hz, 1H), 6.59 (br, 2H), 3.67 (s, 3H), 2.33 (q, J = 7.6 Hz, 2H) , 1.07 (t, J = 7.6 Hz, 311). HRMS (ESI) calculated for C ₂₂ H ₂₁ N ₆ O ₃ [M + H] ⁺ 417.1675, found 417.1678. N- (4- (2- (4-Methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) propionamide (Compound 008)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.15 (s, 1H), 10.08 (br, 1H), 8.85 (s, 1H), 8.04 (s, 1H) , 7.80 (d, J = 8.4 Hz, 2H), 7.35-7.33 (m, 4H), 6.61 (br, 2H), 3.67 (s, 3H), 2, 41 (q, J = 7.6 Hz, 2H), 1.14 (t, J = 7.6 Hz, 3H). HRMS (ESI) calculated for C ₂₂ H ₂₁ N ₆ O ₃ [M + H] ⁺ 417.1675, found 417.1674. 4- (Dimethylamino) -N- (4- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -pentidinyl) -phenyl) -2-butenamide (Compound 009)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.45 (s, 1H), 10.10 (br, 1H), 8.85 (s, 1H), 8.05 (s, 1H) , 7.87 (d, J = 8.8 Hz, 2H), 7.37 (d, J = 8.8 Hz, 2H), 7.30 (br, 2H), 6.82 (td, J = 15.4, 6.0 Hz, 1H), 6.60 (br, 2H), 6.40 (d, J = 15.4 Hz, 1H), 3.63 (s, 3H), 3.27 ( d, J = 5.2 Hz, 2H), 2.33 (s, 6H). HRMS (ESI) calculated for C ₂₅ H ₂₆ N ₇ O ₃ [M + H] ⁺ 472.2097, found 472.2095. 4- (Dimethylamino) -N- (3- (2- (4-methoxyphenylamino) -7-oxo-8 (7H) -puteridinyl) -phenyl) -2-butenamide (Compound 010)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.33 (s, 1H), 10.08 (br, 1H), 8.86 (s, 1H), 8.05 (s, 1H) , 7.83 (d, J 8.0 Hz, 1H), 7.78 (s, 1H), 7.55 (t, J = 8.0 Hz, 1H), 7.32 (br, 2H), 7.11 (d, J = 8.0 Hz, 1H), 6.74 (td, J 15.2, 5.6 Hz, 1H), 6.59 (br, 2H) 6.30 (d, J = 15.2 Hz, 1H), 3.66 (s, 3H), 3.06 (d, J 5.6 Hz, 2H), 2.17 (s, 6H). HRMS (ESI) calculated for C ₂₅ H ₂₄ N ₇ O ₃ [M + H] ⁺ 472.2097, found 472.2094. 4- (2- (4-Methoxyphenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl acrylate (Compound 011)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.15 (s, 1H), 8.87 (s, 1H), 8.06 (s, 11-1), 7.51 (d, J = 8.8 Hz, 2H), 7.45 (d, J = 8.8 Hz, 2H), 7.31 (br, 2H), 6.69 (br, 2H), 6.60 (dd, J = 17.2, 1.6 Hz, 1H), 6.51 (dd, J = 17.2, 9.9 Hz, 1H), 6.22 (dd, J = 9.9, 1.6 Hz , 1H), 3.67 (s, 3H). HRMS (ESI) calculated for C ₂₂ H ₁₈ N ₅ O ₄ [M + H] ⁺ 416.1359, found 416.1359. 4- (Dimethylamino) -N- (4- (7-oxo-2- (phenylamino) -8 (7H) -pteridinyl) phenyl) -2-butenamide (Compound 012)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.37 (s, 11-1), 10.19 (br, 1H), 8.90 (s, 1H), 8.08 (s, 1H), 7.87 (d, J 8.4Hz, 2H), 7.42 (d, J = 7.6Hz, 2H), 7.38 (d, J = 8.4Hz, 2H), 7.03 (br, 1H), 6.88 (t, J = 7.6Hz, 1H), 6.82 (td, J = 15.4, 5.6Hz, 1H), 6.37 (i.e. , J = 15.4 Hz, 1H), 3.14 (d, J = 5.6 Hz, 2H), 2.24 (s, 6H). HRMS (ESI) calculated for C ₂₄ H ₂₄ N ₇ O 2 [M + H] ⁺ 442.1991, found 442.1989. 4- (Dimethylamino) -N- (3- (7-oxo-2- (phenylamino) -8 (7H) -pteridinyl) phenyl) -2-butenamide (Compound 013)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.32 (s, 1H), 10.17 (s, 1H), 8.90 (s, 1H), 8.08 (s, 1H) , 7.81-7.79 (m, 2H), 7.55 (t, J = 8.0 Hz, 1H), 7.41 (d, J = 7.2 Hz, 2H), 7.12 ( d, J = 8.0 Hz, 1H), 7.01 (br, 21-1), 6.87 (t, J = 7.2 Hz, 1H), 6.73 (td, J = 15.2, 5.6 Hz, 1H), 6.28 (d, J = 15.2 Hz, 1H), 3.05 (d, J = 5.6 Hz, 2H ), 2.16 (s, 6H). HRMS (ESI) calculated for C ₂₄ H ₂₄ N ₇ O ₂ [M + H] ⁺ 442.1991, found 442.1996. N- (4- (7-Oxo-2- (phenylamino) -8 (7H) -pteridinyl) phenyl) acrylamide (Compound 014)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.44 (s, 1H), 10.19 (br, 1H), 8.90 (s, 1H), 8.09 (s, 1H) , 7.88 (d, J = 8.4Hz, 2H), 7.41-7.38 (m, 4H), 7.03 (br, 2H), 6.88 (t, J = 7.2 Hz, 1H), 6.53 (dd, J = 16.8, 10.4 Hz, 1H), 6.35 (dd, J = 16.8, 1.6 Hz, 1H), 5.84 (dd , J = 10.4, 1.6 Hz, 1H). HRMS (ESI) calculated for C ₂₁ H ₁₇ N ₆ O ₂ [M + H] ⁺ 385.1413, found 385.1405. N- (3- (7-Oxo-2- (phenylamino) -8 (7H) -pteridinyl) phenyl) acrylamide (Compound 015)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.42 (s, 1H), 10.19 (s, 1H), 8.91 (s, 1H), 8.09 (s, 1H) , 7.84-7.81 (m, 2H), 7.57 (t, J = 8.0 Hz, 1H), 7.41 (br, 2H), 7.15 (d, J = 7.6 Hz, 1H), 7.02 (br, 2H), 6.87 (t, J = 7.6 Hz, 1H), 6.45 (dd, J = 16.8, 10.4 Hz, 1H), 6.26 (dd, J = 16.8, 1.6 Hz, 1H), 5.77 (dd, J = 10.4, 1.6 Hz, 1H). HRMS (ESI) calculated for C ₂₁ H ₁₇ N ₆ O ₂ [M + H] ⁺ 385.1413, found 385.1413. N- (4- (2- (4-Chloro-phenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) -acrylamide (Compound 016)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.46 (s, 1H), 10.34 (s, 1H), 8.92 (s, 1H), 8.11 (5, 1H) , 7.88 (d, J = 8.8 Hz, 2H), 7.41-7.36 (m, 4H), 7.06 (br, 2H), 6.53 (dd, J = 16.8 , 10.4 Hz, 1H), 6.36 (dd, J = 16.8, 1.6 Hz, 1H), 5.84 (dd, J = 10.4, 1.6 Hz, 1H). HRMS (ESI) calculated for C ₂₁ H ₁₆ N ₆ O ₂ Cl [M + H] ⁺ 419.1023, found 419.1031. N- (3- (2- (4-Chloro-phenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) -acrylamide (Compound 017)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.44 (s, 1H), 10.34 (br, 1H), 8.93 (s, 1H), 8.11 (s, III) , 7.84 (s, III), 7.81 (d, J = 8.4 Hz, 11-I), 7.59 (t, J = 8.0 Hz, 1H), 7.43 (d, J = 7.2 Hz, 2H), 7.15 (d, J 7.6 Hz, 1H), 6.46 (dd, J = 16.8, 10.4 Hz, 1H), 6.26 (dd , J = 16.8, 1.8 Hz, 1H), 5.77 (dd, J 10.12, 1.8 Hz, 1H). HRMS (ESI) calculated for C ₂₁ H ₁₆ N ₆ O ₂ Cl [M + H] ⁺ 419.1023, found 419.1027. N- (3- (2- (4-morpholinophenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) -acrylamide (Compound 018)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.43 (s, 1H), 10.06 (s, 1H), 8.84 (s, 1H), 8.03 (s, 1H) , 7.92 (br, 1H), 7.72 (s, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.27 (br, 2H), 7.12 (d, J = 7.2 Hz, 1H), 6.58 (br, 2H), 6.45 (dd, J = 16.8, 10.4 Hz, 1H), 6.26 (d, J = 16.8 Hz, 1H), 5.78 (d, J = 10.4 Hz, 1H), 3.71 (br, 4H), 2.94 (br, 4H). HRMS (ESI) calculated for C ₂₅ H ₂₄ N ₇ O ₃ [M + H] ⁺ 470.1941, found 470.1939. N- (4- (2- (4- (4-Methyl-1-piperazinyl) phenylamino) -7-oxo-8 (7H) -puteridinyl) -phenyl) acrylamide (Compound 019)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.51 (s, 1H), 10.06 (s, 11-1), 8.83 (s, 1H), 8.03 (s, 1H), 7.89 (d, J = 8.4Hz, 2H), 7.37 (d, J = 8.4Hz, 2H), 7.17 (d, J = 6.4Hz, 1H) , 6.56-6.49 (m, 3H), 6.34 (d, J = 16.8Hz, 1H), 5.85 (d, J = 10.8Hz, 1H), 2.94 ( br, 4H), 2.37 (br, 4H), 2.20 (s, 3H). HRMS (ESI) calculated for C ₂₆ H ₂₇ N ₈ O ₂ [M + H] ⁺ 483.2257, found 483.2259. N- (3- (2- (4- (4-Methyl-1-piperazinyl) -phenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) acrylamide (Compound 020)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.45 (s, 1H), 10.06 (s, 1H), 8.84 (s, 1H), 8.04 (s, 1H) , 7.93 (br, 1H), 7.73 (s, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.25 (br, 2H), 7.12 (d, J = 8.0 Hz, 1H), 6.57 (br, 2H), 6.46 (dd, J = 16.8, 10.4 Hz, 111), 6.27 (dd, J = 16.8 , 1.8 Hz, 1H), 5.78 (dd, J = 10.4, 1.8 Hz, 1H), 2.98 (br, 4H), 2.42 (br, 4H), 2.22 (s, 3H). HRMS (ESI) calculated for C ₂₆ H ₂₇ N ₈ O ₂ [M + H] ⁺ 483.2257, found 483.2259. N- (3- (7-Oxo-2- (4- (1-piperidinyl) phenylamino) -8 (7H) -pteridinyl) phenyl) acrylamide (Compound 021)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.44 (s, 1H), 10.03 (s, 1H), 8.83 (s, 1H), 8.02 (s, 1H) , 7.94 (br, 1H), 7.73 (s, 1H), 7.55 (t, J = 8.0 Hz, 111), 7.24 (br, 2H), 7.11 (d, J 8.0 Hz, 1H), 6.57 (br, 2H), 6.46 (dd, J = 17.0, 10.2 Hz, 1H), 6.26 (dd, J = 17.0, 1.8 Hz, 1H), 5.77 (dd, J = 10.2, 1.8 Hz, 1H), 2.95 (br, 4H), 1.57 (br, 4H), 1.49 ( br, 2H). HRMS (ESI) calculated for C ₂₆ H ₂₆ N ₇ O ₂ [M + H] ⁺ 468.2148, found 468.2146. N- (3- (7-Oxo-2- (4- (1-pyrrolidinyl) phenylamino) -8 (7H) -pteridinyl) phenyl) acrylamide (Compound 022)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.40 (s, 1H), 9.92 (s, 1H), 8.79 (s, 1H), 7.99 (s, HI) , 7.90 (br, 1H), 7.74 (br, 1H), 7.54 (t, J = 8.0 Hz, 1H), 7.20 (br, 2H), 7.10 (d, J = 8.0 Hz, 1H), 6.46 (dd, J = 17.0, 10.2 Hz, 1H), 6.26 (dd, J = 17.0, 1.8 Hz, 1H), 6.20 (br, 2H), 5.77 (dd, J = 10.2, 1.8Hz, 1H), 3.10 (br, 4H), 1.91 (br, 4H). HRMS (ESI) calculated for C ₂₅ H ₂₄ N ₇ O ₂ [M + H] ⁺ 454.1991, found 454.1995. N- (3- (2- (4- (Diethylamino) phenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (Compound 023)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.42 (s, 1H), 9.92 (s, 1H), 8.80 (s, 11-1), 8.00 (s, 1H), 7.92 (br, 1H), 7.73 (s, 1H), 7.53 (t, J = 8.0Hz, 1H), 7.19 (br, 2H), 7.09 ( d, J = 8.0 Hz, 1H), 6.46 (dd, J = 17.0, 10.2 Hz, 1H), 6.32 (br, 2H), 6.27 (dd, J = 17 , 0, 1.8 Hz, 1H), 5.76 (dd, J = 10.2, 1.8 Hz, 1H), 3.20 (br, 4H), 1.00 (t, J = 6, 8 Hz, 6H). HRMS (ESI) calculated for C ₂₅ H ₂₆ N ₇ O ₂ [M + H] ⁺ 456.2148, found 456.2143. N- (3- (2- (4- (Acetylamino) phenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (Compound 024)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.43 (s, 1H), 10.16 (br, 1H), 9.78 (s, 1H), 8.87 (s, III) , 8.06 (s, 114), 7.82 (d, J = 8.0 Hz, 1H), 7.79 (s, 1H), 7.56 (t, J = 8.0 Hz, 1H) , 7.32 (br, 21-1), 7.23 (br, 21-1), 7.15 (d, J = 8.0 Hz, 1H), 6.46 (dd, J = 17.0 , 10.2 Hz, 1H), 6.25 (dd, J = 17.0, 1.8 Hz, 1H), 5.76 (dd, J = 10.2, 1.8 Hz, 1H), 1 , 98 (s, 3H). HRMS (ESI) calculated for C ₂₃ H ₂₀ N ₇ O ₃ [M + H] ⁺ 442.1628, found 442.1624. 4- (8- (3-acrylamidophenyl) -7-oxo-7,8-dihydropteridinyl-2-amino) benzamide (Compound 025)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.43 (s, 1H), 10.40 (s, 1H), 8.95 (s, 1H), 8.13 (s, 1H) , 7.86 (s, 1H), 7.79 (d, J = 8.0 Hz, 1H), 7.71 (br, 1H), 7.61 (t, J 8.0 Hz, 1H), 7.55 (d, J = 7.6 Hz, 2H), 7.47 (br, 2H), 7.18 (d, J = 7.6 Hz, 2H), 6.44 (dd, J = 17 , 0, 10.2 Hz, 1H), 6.25 (dd, J = 17.0, 1.8 Hz, 1H), 5.76 (dd, J = 10.2, 1.8 Hz, 1H) , HRMS (ESI) calculated for C ₂₂ H ₁₈ N ₇ O ₃ [M + H] ⁺ 428.1471, found 428.1476. N- (3- (2- (4-methoxyphenylamino) -6-methyl-7-oxo-8 (7H) -pteridinyl) -phenyl) -acrylamide (Compound 026)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.42 (s, 1H), 9.93 (br, 1H), 8.78 (s, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.77 (s, 11-1), 7.56 (t, J = 8.0 Hz, 1H), 7.31 (br, 2H), 7.11 (d, J = 8.0 Hz, 1H), 6.58 (br, 2H), 6.45 (dd, J = 17.0, 10.2 Hz, 1H), 6.26 (d, J r 17.0 Hz, 11-1), 5.77 (d, J = 10.2 Hz, 1H), 3.65 (s, 3H), 2.42 (s, 3H). HRMS (ESI) calculated for C ₂₃ H ₂₁ N ₆ O ₃ [M + H] ⁺ 429.1675, found 429.1675. N- (3- (8- (4-methoxyphenyl) -7-oxo-7,8-dihydropteridine-2-amino) phenyl) acrylamide (Compound 027)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.17 (s, 1H), 10.01 (s, 1H), 8.89 (s, 1H), 8.07 (s, 1H) , 7.63 (br, 1H), 7.33 (d, J = 8.8 Hz, 2H), 7.27 (d, J = 8.0 Hz, 1H), 7.23 (d, J = 8.0 Hz, 1H), 7.11 (d, J = 8.8 Hz, 2H), 6.89 (br, 1H), 6.46 (dd, J = 17.0, 10.2 Hz, 1H), 6.25 (dd, J = 17.0, 1.8 Hz, 1H), 5.74 (dd, J = 10.2, 1.8 Hz, 1H), 3.85 (s, 3H ). HRMS (ESI) calculated for C ₂₂ H ₁₉ N ₆ O ₃ [M + H] ⁺ 415.1519, found 415.1519. 2- (3-aminophenylamino) -8- (4-methoxyphenyl) -7 (8H) -pteridinone (Compound 028)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 9.91 (s, 1H), 8.85 (s, 1H), 8.04 (s, 1H), 7.35 (d, J 8 , 8Hz, 2H), 7.16 (d, J = 8.8Hz, 2H), 6.68-6.65 (m, 3H), 6.16 (d, J = 7.2Hz, 1H ), 4.63 (s, 2H), 3.86 (s, 3H). HRMS (ESI) calculated for C ₁₉ H ₁₇ N ₆ O ₂ [M + H] ⁺ 361.1413, found 361.1411. N- (4- (8- (4-Methoxyphenyl) -7-oxo-7,8-dihydropteridine-2-amino) phenyl) acrylamide (Compound 029)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.19 (br, 1H), 10.03 (s, 1H), 8.87 (s, 1H), 8.05 (s, 1H) , 7.36-7.34 (m, 6H), 7.16 (d, J = 8.4 Hz, 2H), 6.41 (dd, J = 17.0, 10.2 Hz, 1H), 6.23 (dd, J = 17.0, 1.6 Hz, 1H), 5.72 (dd, J = 10.2, 1.6 Hz, 1H), 3.92 (s, 1H). HRMS (ESI) calculated for C ₂₂ H ₁₉ N ₆ O ₃ [M + H] ⁺ 415.1519, found 415.1524. 2- (4-aminophenylamino) -8- (4-methoxyphenyl) -7 (8H) -pteridinone (Compound 030)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 9.87 (s, 1H), 8.77 (s, 1H), 7.97 (s, 1H), 7.32 (d, J = 8.8 Hz, 21-I), 7.13 (d, J = 8.8 Hz, 2H), 7.08 (br, 2H), 6.24 (br, 2H), 4.84 (s, 2H), 3.88 (s, 3H). HRMS (ESI) calculated for C ₁₉ H ₁₇ N ₆ O ₂ [M + H] + 361.1413, found 361.1417. N- (4- (2- (2-Methoxy) -4- (4-methoxy-1-piperazinyl) phenylamino) -7-oxo-8 (7H) -pteridinyl) phenyl) acrylamide (Compound 031)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.43 (s, 1H), 8.80 (s, 1H), 8.42 (s, 1H), 8.03 (s, 1H) , 7.85 (d, J = 8.6 Hz, 2H), 7.34 (d, J = 8.6 Hz, 2H), 7.25 (d, J = 8.8 Hz, 1H), 6 , 54-6.48 (m, 21-1), 6.33 (dd, J = 17.0, 1.6 Hz, 1H), 6.02 (br, 1H), 5.84 (dd, J = 10.2, 1.6Hz, 1H), 3.76 (s, 31-1), 3.02 (br, 4H), 2 , 43 (br, 4H), 2.23 (s, 3H). HRMS (ESI) calculated for C ₂₇ H ₂₉ N ₈ O ₃ [M + FI] ⁺ 513.2363, found 513.2362. N- (3- (2- (2-Methoxy-4- (4-methyl-1-piperazinyl) -phenylamino) -7-oxo-8 (7H) -pteridinyl) -phenyl) -acrylamide (Compound 032)

¹ H-NMR (400 MHz, DMSO-d ₆ ): δ 10.41 (s, 1H), 8.80 (s, 1H), 8.44 (br, 1H), 8.0 2 (s, 1H), 7.86 (br, 1H), 7.71 (s, 1H), 7.52 (t, J = 8.0 Hz, 1H), 7.30 (d, J = 7.6 Hz, 1H), 7.09 (d, J = 8.0 Hz, 1H), 6.53 (s, 1H), 6.46 (dd, J = 17.0, 10.2 Hz, 1H), 6.26 (dd , J = 17.0, 1.8 Hz, 1H), 6.02 (br, 1H), 5.78 (dd, J = 10.2, 1.8 Hz, 1H), 3.76 (s, 3H), 3.04 (br, 4H), 2.44 (br, 4H), 2.23 (s, 3H). HRMS (ESI) Calculated for C ₂₇ H ₂₉ N ₈ O ₃ [M + H] ⁺ 513.2363, found 513.2361.

Example 2

Bioactivity Assay - 1

The inhibitory effects of the compounds provided in the present invention on EGFR kinase activity in vitro were tested as follows:
In vitro enzyme activity assay: wild-type and various EGFR mutants (T790M, L858R, L861Q, L858R / T790M) and a Z'-Lyte kinase assay kit were purchased from Invitrogen. Ten concentration gradients of 5.1 x 10 ^-11 mol / l to 1.0 x 10 ^-6 mol / l were adjusted for all compounds to be tested.

Concentrations of various kinases were determined by optimization of the experiment and the respective concentrations were: EGFR (PV3872, Invitrogen) 0.287 μg / μl, EGFR-T790M (PV4803, Invitrogen) 0.174 μg / μl, EGFR-L858R (PV4128, Invitrogen) 0.054 μg / μl, EGFR-L858R / T790M (PV4879, Invitrogen) 0.055 μg / μl.

Compounds were diluted in DMSO from 5.1 x 10 ^-9 M to 1 x 10 ^-4 M to one-third. In 96 μl of water, 4 μl of compound was dissolved, resulting in a 4-fold solution of the compound. 40 μM ATP was dissolved in 1.33 x kinase buffer and a kinase / peptide mixture comprising 2 x kinase, 4 μM tyrosine and four peptides was prepared for use. 10 μl kinase reaction system included 2.5 μl compound solution, 5 μl kinase / peptide mixture and 2.5 μl ATP solution. Instead of the kinase / peptide mixture, 5 μl phosphopeptide solution was used as 100% phosphorylation control. 2.5 μl of 1.33 x kinase buffer was used to replace the 100% inhibitor control ATP solution and 2.5 μl of 4% DMSO solution was used to replace compound solution as 0% inhibitor control. After thoroughly mixing the solution within the plate, the plate was incubated for 1.5 hours at room temperature. Into each well, 5 μl of DevelopmentSolution was added, and then the plate was incubated for 1 more hour at room temperature and within this time non-phosphorylated peptide was cleaved. Finally, the reaction was stopped by adding 5 μl of Stop Reagent. The plate was measured with an EnVision Multilabel Reader (Perkin Elmer). The experimental data was calculated using GraphPad Prism Version 4.0. Each experiment was repeated three times.

The test results are shown in Table 1. Table 1 Connection no. Inhibitory effect on EGFR kinase (IC ₅₀ , nM) T790M WT L858R T790M / L858R 001 > 10000 > 10000 > 10000 > 10000 002 665 446 546 606 003 8698 8011 4082 3297 004 > 10000 3181 4877 > 10000 005 > 10000 > 10000 87159 > 10000 006 19.4 10.6 10.1 8.4 007 > 10000 > 10000 > 10000 > 10000 008 > 10000 > 10000 > 10000 > 10000 009 10.9 60.7 81.11 39.1 010 2253 8086 6084 2022 011 67 84.8 70.3 012 5980 5342 1920 013 86.6 101 39.5 014 16.1 26.4 113.2 015 14.7 12.4 5.28 016 7580 115.9 1491 017 67 84.8 70.3 018 7.94 5.83 3.02 019 1260 6382 738 020 1.47 1.2 0.824 021 19.7 11.7 5.49 022 23.3 12.6 5.33 023 23.1 16.3 5.57 024 15.8 13.4 5.12 025 12.7 9.64 4.39 026 7.24 5.93 16 027 2000 1710 1200 028 10603 18881 3534 029 > 10000 1318 > 10000 030 3415 23758 6751 031 429 376 229 032 3.67 2.36 1.17 WZ4002 9.58 2.6 1.02

Bioactivity Assay - 2

Cell proliferation and growth inhibition analysis: H1975 cells (non-small cell lung cancer cells, EGFR ^{L858R / T790M} ), HCC827 (non-small cell lung cancer cells, EGFR ^{del E746-A750} ), A549 (non-small cell lung cancer cells, EGFR wild type), BT474 (breast cancer cells, Her2 overexpression), SK-BR-3 (breast cancer cells, Her2 overexpression), MCF-7 (breast cancer cells, Her2 overexpression) were obtained from the ATCC. Cell proliferation was assessed by an MTS assay. The cells were subjected to the process conditions for 72 hours, and the number of cells for each cell line used in the experiment was adjusted according to the absorbance value (the absorbance value at 490 nm was 1.3-2.2). For the compounds to be tested, 6 concentration gradients (0.1 nM to 10 μM) were set up with six parallel controls for each concentration.

Cells of the type H1975, HCC827, A549, BT474, MCF-7 and SK-BR-3 were cultured in the appropriate medium. After recovery, the cells were re-culture at least twice and then used in experiments. Cells in the log phase were trypsinized and in the medium resuspended. H11975 (1000 cells per well), BT474 (1500 cells per well), MCF-7 (1500 cells per well), HCC827 (2000 cells per well), SK-BR-3 (2000 cells per well), A549 (2000 cells per well) were seeded in 96-well plates, the volume was 100 μl, and 6 rows and 7 columns were occupied. The plate was placed in an incubator at 37 ° C and 5% carbon dioxide overnight. The compounds were dissolved in DMSO to give a concentration of 10 μM and then gradually diluted to give a compound concentration of 10 μM, 1 μM, 0.1 μM, 0.01 μM, 0.001 μM, 0 , 0001 μM received. 2 μl of compound solution was added to 998 μl of medium and the resulting mixture was mixed thoroughly. 100 μl of mixture was added to a 96-well plate. 2 μl of DMSO was used as 0% inhibition control to replace the compound solution. After the cells had been cultured for 68 hours, 20 μl MTT (5 mg / ml) was added. After 4 hours, the supernatant was discarded and 150 μl of DMSO was added. The resulting system was shaken for 10 minutes and the plate was read with a Synergy HT (Bio TeK) (OD490). Data were calculated using GraphPad Prism Version 4.0 and a nonlinear regression model using a dose-response curve yielded the IC ₅₀ value.

The test results are shown in Tables 2 and 3. Table 2 Connection no. Inhibitory effect on cell proliferation (IC ₅₀ , μM) HCC827 H1975 001 > 10 > 10 002 1.29 2.75 003 3.01 > 10 004 3.33 > 10 005 > 10 > 10 006 0.009 0,133 007 > 10 > 10 008 > 10 > 10 009 0.91 4.65 010 4.16 > 10 011 2.29 5.6 012 4.42 18.9 013 0.914 3.51 014 68.1 77.8 015 0,015 0.437 016 68.7 20.8 017 0.163 0.82 018 0,017 0,216 019 0.676 8.87 020 0,002 0.043 021 0.02 0,238 022 0.073 0.531 023 0.031 0.477 024 0.229 6.08 025 0.604 7.3 026 0,013 1.22 027 0.408 3.05 028 15.9 82.8 029 0.543 0.953 030 4.65 15.3 031 0.466 2.88 032 0,004 0,038 WZ4002 0,014 0,039 Iressa 0,006 13 Table 3 Connection no. Inhibitory effect on cell proliferation (IC ₅₀ , μM) A549 SK-BR-3 MCF-7 BT474 001 > 10 > 10 > 10 > 10 002 1.23 1.57 1.51 > 10 003 > 10 > 10 > 10 > 10 004 > 10 > 10 > 10 > 10 005 > 10 > 10 > 10 > 10 006 0.73 2.35 15.0 2.24 007 > 10 > 10 > 10 > 10 008 6.12 > 10 > 10 > 10 009 2.24 1.15 2.41 0.84 010 > 10 7.17 3.60 3.43

Bioactivity Assay - 3

Kinase selectivity analysis: A kinase selectivity experiment was performed by Shanghai ChemPartner using a Caliper assay screening platform. All kinases and other materials were sourced from commercial companies. Staurosporine and PI103 were used as control compounds in testing the inhibitory activity of the compound on various kinases.

I. Mobility shift assay

• 1. Preparation of Kinase Matrix Buffer and Kinase Test Stop Buffer: 1) 1X Kinase Matrix Buffer: 50 mM HEPES pH 7.5, 0.0015% Brij-35, 10 mM MgCl ₂ , 2 mM DTT; 2) Stop buffer: 100 mM HEPES, pH 7.5, 0.015% Brij-35, 0.2% Coating Reagent # 3, 50 mM EDTA.
• 2. Preparation of a compound solution: 1) The compound was diluted in 100% DMSO to a concentration 50 times the highest concentration used in the test, and to each well of a 96-well plate was added 100 μl of the above compound solution , 2) 30 μl of compound solution was transferred into 60 μl of 100% DMSO in the adjacent well, and the above procedure was repeated to prepare a dilution series of the compound. 3) 100 μl of 100% DMSO was added to two empty wells as the no-link and no-enzyme controls, and the plate was labeled as the stock solution plate. 4) 10 μl of solution from the stock solution plate was placed in another 96-well plate as a temporary plate, and 90 μl of 1 × kinase buffer was placed in each well, the temporary plate was placed on a shaker to uniformly mix the compound solution.
• 3. Preparation of the assay plate: 5 μl of solution from each well of the 96-well temporary plate was placed in a 384 well plate and the experiment was repeated independently.
• 4. Kinase reaction: 1) Prepare a 2.5X enzyme solution by adding the kinase to 1X kinase matrix buffer. 2) Preparation of a 2.5X peptide solution: FAM-labeled peptide and ATP were added to 1X kinase matrix buffer. 3) 2.5x enzyme solution was transferred to the assay plate. 4) The assay plate comprised 5 μl of the compound solution in 10% DMSO. 5) Into each well of the 384-well assay plate was added 10 μl of the 2.5X enzyme solution. 6) The assay plate was incubated for 10 minutes at room temperature. 7) 2.5X peptide solution was transferred to the assay plate and placed in each well of the 384 wells. Assay plate was given 10 μl of peptide solution. 8) Kinase reaction and termination: After incubation at 28 ° C for a while, 25 μl stop buffer was added to stop the reaction.
• 5. Caliper readings: The experimental data were collected on Caliper.
• 6. Curve Fitting: 1) Conversion data copied from the Caliper program. 2) The conversion value was converted into the inhibition rate,% inhibition rate = (maximum conversion value) / (maximum - minimum) × 100, where "maximum" stands for the DMSO control and "minimum" stands for the low control.

II. Kinase-Glo analysis

• 1. Preparation of 1X Kinase Buffer for the PI3Ka Kinase Assay: 1X Kinase Buffer: 50mM HEPES, pH 7.5, 3mM MgCl ₂ , 1mM EGTA, 100mM NaCl, 0.03% CHAPS, 2mM DTT.
• 2. Preparation of compounds for PI3Ka kinase assay: 1) Serial dilution of the compound and preparation of stock plate. The compound was diluted in 100% DMSO to a concentration 100 times the highest concentration used in the reaction, and to each well of a 96-well plate was added 100 μl of the above compound solution. Thirty μl of the compound solution was transferred into 60 μl of 100% DMSO in the adjacent well, and the above procedure was repeated to make a dilution series of the compound. 100 μl of 100% DMSO was added to two empty wells as no-link and no-enzyme controls, and the plate labeled as stock plate. 2) Preparation of the temporary plate. 4 μl of solution from the stock solution plate was placed in another 96-well plate, and each well was charged with 96 μl of 1 × kinase buffer, and the temporary plate was placed on a shaker and shaken for 10 minutes to uniformly mix the compound solution , 3) Preparation of assay plate: 2.5 μl of solution from each well of the 96-well temporary plate was placed in a 384-well plate and the experiment was repeated independently.
• 3. Kinase PI3Ka reaction: 1) Preparation of a 4X kinase solution. PI3Ka solution was prepared in 1 × kinase buffer whose concentration was 4 times the final concentration in the assay. Into each well of the assay plate (except the control well into which 2.5 μl of kinase buffer was added) was added 2.5 μl of 1 × kinase solution given, and the assay plate was shaken. 2) Preparation of a 2 × substrate solution. A PIP2 substrate ATP solution was prepared in 1 × kinase reaction buffer whose concentration was twice the final concentration in the assay. Into each well of the assay plate was added 5 μl of substrate solution and shaken to mix the contents evenly. 3) The kinase reaction was carried out by incubation at room temperature for 1 hour.
• 4. kinase assay. Kinase-Glo reagent was brought to RT, and 10 μl of Kinase-Glo reagent was added to the assay plate to stop the reaction. After simple mixing, the plate was centrifuged and shaken slowly on an oscillator for 15 minutes, and then the data was read on a luminescence reader.
• 5. The data was read and collected on Flexstation.
• 6. Curve Fitting: RLU data has been copied from the Flex Station program. The data were converted to% inhibition rate. % Inhibition rate = (RLU of the sample - minimum) / (maximum - minimum) x 100, where "maximum" stands for the RLU data of the no enzyme control and "minimum" stands for the RLU value of the DMSO control group.

III. BRAF analysis

• 1. Preparation of 1 × kinase buffer: 50 mM HEPES, pH 7.5, 10 mM MgCl ₂ , 1 mM EGTA, 0.01% BRIJ-35.
• 2. Preparation of Compounds for BRAF Kinase Assay: 1) Serial dilution of the compound and preparation of a stock solution plate. The compound was diluted in 100% DMSO to a concentration 100 times the highest concentration used in the reaction, and to the wells of a 96-well plate was added 100 μl of the above compound solution. Thirty μl of the compound solution was transferred into 60 μl of 100% DMSO in the adjacent well, and the above procedure was repeated to make a dilution series of the compound. 100 μl of 100% DMSO was added to two empty wells as the no-link and no-enzyme controls, and the plate was labeled as the stock solution plate. 2) Preparation of the temporary plate. 4 μl of compound solution from the stock solution plate was transferred to another 96-well plate, 96 μl of 1 × kinase buffer was added to each well, and the temporary plate was placed on a shaker and shaken for 10 minutes to uniformly mix the compound. 3) Preparation of assay plate: 2.5 μl of solution from each well of the 96-well temporary plate was placed in a 384-well plate and the experiment was repeated independently.
• 3. Kinase BRAF reaction: 1) Preparation of a 2 × kinase solution. BRAF solution was prepared in 1 × kinase buffer whose concentration was twice the final concentration in the assay. Into each well of the assay plate (except the control well into which 5 μl of kinase buffer was added) was added 5 μl of kinase solution and the assay plate was shaken. 2) Preparation of a 4 × substrate solution. A fluorescein MAP2K1-ATP substrate solution was prepared in 1 × kinase buffer whose concentration was 4 times the final concentration in the assay. Into each well of the assay plate was added 2.5 μl of substrate solution to start the reaction and the assay plate was shaken. 3) The kinase reaction was carried out by incubation at room temperature for 1 hour.
• 4. kinase assay. A test solution was prepared in antibody dilution buffer whose concentration was twice the following final concentration: 2 nM antibody, 10 μM EDTA. Into each well of the assay plate was added 10 μl of test solution to stop the assay. After simple mixing, the plate was centrifuged and incubated for at least 30 minutes.
• 5. Data Reading: The data were read on Envision (excitation at 340 nm, emission at 520 nm, 495 nm).
• 6. Curve Fitting: RLU data has been copied by the Envision program. The rate of RFU of 520 nm / 495 nm was calculated and converted to% inhibition rate. % Inhibition rate = (maximum rate of the sample) / (maximum - minimum) x 100, where "maximum" stands for the DMSO control rate and "minimum" stands for the no enzyme control rate. Curve fitting was done using XLFit Excel Add-in version 4.3.1.

The test results are shown in the following table. Table 4 kinase % Inhibition (10 μM) % Inhibition (1 μM) Connection 020 Connection 032 Connection 020 Connection 032 HER2 98 100 97 89 HER4 72 77 60 60 FLT1 58 11 17 9.4 FLT3 99 91 82 60 CDK2 49 48 18 25 BLK 100 99 99 85 JAK2 36 22 22 9.9 LCK 73 54 18 14 LYNA 40 20 11 6.1 cKit -3.2 -4.8 -12 -9.3 PIM1 54 8.9 8.2 2.1 FGFR3 33 13 14 11 FGFR1 63 42 19 7.9 PDGFRA 35 18 4.9 1.2 PDGFRB 84 61 42 14 KDR 73 29 25 3.8 SRC 72 33 27 6.6 ABL 39 13 11 5.8 AUR B 87 69 52 32 C-MET 53 28 27 13 BRAF 21 -5.2 -7.0 0.35 PKACa 4.7 4.0 15 6.9 IKKB 2.0 -4.3 9.4 -3.3 IGF1R 36 37 21 15 GSK3b 34 16 16 12 P38a 21 1.2 2.8 -5.6 ERK1 19 -4.0 1.3 11

Bioactivity Assay - 4

The inhibitory effects of the compounds provided in the present invention on BLK and FLT3 kinase activity in vitro were tested as follows, wherein BLK, FLT3 were obtained from BPS and stauroporin was used as a control compound:
Preparation of 1 × kinase matrix buffer and stop buffer. 1 × kinase matrix buffer: 50 mM HEPES, pH 7.5, 0.0015% Brij-35, 10 mM MgCl ₂ , 2 mM DTT; Stop buffer: 100mM HEPES, pH 7.5, 0.0015% Brij-35, 0.2% Coating Reagent # 3, 50mM EDTA.

Preparation of a compound solution: 1) The compound was diluted in 100% DMSO to a concentration 50 times the highest concentration used in the test. In wells of a 96-well plate, 100 μl of the above compound solution was added. From the above compound solution, a dilution series was prepared to the last desired concentration. Into two empty wells of the same 96-well plate was added 100 μl of 100% DMSO as a no-link and no-enzyme control, and this plate was used as an original plate.

Making the temporary plate. 10 μl of compound solution was transferred from the original plate to another 96-well plate as a temporary plate; into each well of the temporary plate was added 90 μl of 1 × kinase buffer and the temporary plate was shaken for 10 minutes.

Preparation of Assay Plate: 5 μl of solution from each well of the temporary plate was placed in a 384 well plate and repeated controls were set.

Kinase reaction: 2.5 x kinase solution and 2.5 x peptide solution were prepared. 2.5 x kinase solution was transferred to the assay plate. The assay plate comprised 5 μl of the compound solution in 10% DMSO. Into each well of the 384-well assay plate was added 10 μl of the 2.5 x kinase solution. The plate was incubated for 10 minutes at room temperature. Into each well was added 2.5X peptide solution. After incubation at 28 ° C for a while, 25 μl of stop buffer was added to stop the reaction. The experimental data were collected on Caliper. A curve has been adjusted. The experimental data was copied from the Caliper program and converted into the inhibition rate. % Inhibition rate = (maximum conversion value) / (maximum - minimum) × 100, where "maximum" stands for the DMSO control and "minimum" stands for a low control.

The test results are shown in the following Table 5. Table 5 connection Inhibitory effect on the kinase (IC ₅₀ , nM) BLK FLT3 001 > 10000 27 002 3037 128 003 4168 137 004 > 10000 206 005 > 10000 30 006 69 1591 007 > 10000 1486 008 > 10000 115 009 5312 152 010 106 4387 011 89 48 012 6321 160 013 171 3778 014 7479 129 015 124 1859 016 > 10000 358 017 156 2,825 018 41 798 019 796 51 020 <14 322 021 83 878 022 98 1084 023 83 1757 024 148 3548 025 76 1791 026 96 568 027 5656 119 028 7946 34 029 > 10000 100 030 > 10000 43 031 812 180 032 463 1169 staurosporine 5.6 0.28

The present invention is illustrated by the specific examples. It should be understood, however, that the scope of the invention should not be limited to these specific examples, but is defined in the claims, and any equivalent variation of the invention should be within the scope of the invention.

Claims (10)

Compound having the structure of general formula I:

wherein A and B are benzene rings or five- or six-membered heterocycles having different substituents; C is selected from one of the following groups:

wherein X is selected from O, S and Se, R ^{1 is} hydrogen, a halogen atom, C ₁ -C ₆ alkoxy, an optionally substituted C ₁ -C ₆ alkyl, an optionally substituted aryl or an optionally substituted aralkyl; R ^{2 are} each independently selected from hydrogen, a halogen, a C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, sulfonate group, sulfamoyl, carbamoyl, carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl is selected; R ^{3 are} each independently selected from hydrogen, halo, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, sulfonate group, sulfamoyl, carbamoyl, Carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl; R _a and R _{b are} independently selected from an alkyl and an alkenyl; and m and n are independently selected from 0, 1, 2, 3 or 4. A compound according to claim 1, wherein the compound has the structure of general formula II:

wherein Y is selected from N, CH; Z is selected from N, CR ⁶ ; R ^{1 is} hydrogen, halogen, C ₁ -C ₆ alkoxy, an optionally substituted C ₁ -C ₆ alkyl, an optionally substituted aryl or an optionally substituted aralkyl; R ^{3 is} independently selected from hydrogen, amino, hydroxy, optionally substituted acyloxy, alkoxy, halogen, optionally substituted alkyl, CN, a sulfonate group, sulfamoyl, carboxy, morpholinyl, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b optionally substituted acylamino, optionally substituted alkoxyformyl and carbamoyl; R ⁴ , R ⁵ , R ⁶ and R ^{7 are} each independently selected from hydrogen, halo, C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, a sulfonate group, sulfamoyl, carbamoyl, carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl are selected; R _a and R _{b are selected} from an alkyl and an alkenyl; and n and m each = 0, 1, 2, 3 or 4. A compound according to claim 1, wherein the compound has the structure of general formula III:

wherein R ^{1 is} hydrogen, halogen, C ₁ -C ₆ alkoxy, an optionally substituted C ₁ -C ₆ alkyl, an optionally substituted aryl or an optionally substituted aralkyl; R ^{3 is} independently selected from hydrogen, amino, hydroxy, optionally substituted acyloxy, alkoxy, halogen, optionally substituted alkyl, CN, a sulfonate group, sulfamoyl, carboxy, morpholinyl, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b optionally substituted acylamino, optionally substituted alkoxyformyl and carbamoyl; R ⁵ , R ⁶ and R ^{7 are} each independently selected from hydrogen, a halogen, a C ₁ -C ₆ alkoxy, hydroxy, optionally substituted acyloxy, amino, optionally substituted acylamino, optionally substituted C ₁ -C ₆ alkyl, CN, one Sulfonate group, sulfamoyl, carbamoyl, carboxy, optionally substituted alkoxyformyl, optionally substituted phenyl, optionally substituted N-alkylpiperazinyl, optionally substituted morpholinyl, optionally substituted piperidinyl, optionally substituted pyrrolyl, optionally substituted pyrrolidinyl, -NR _a R _b , optionally substituted pyridyl; R _a and R _{b are selected} from an alkyl and an alkenyl; and m and n each = 0, 1, 2, 3 or 4. A compound according to claim 3, wherein R ^{1 is selected} from H and a C ₁ -C ₆ alkyl; R ^{3 is} hydrogen, amino, hydroxy, acyloxy, alkoxy, halogen, hydroxy, alkyl, CN, a sulfonate group, sulfamoyl, carboxy, morpholinyl, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b , acylamino and Carbamoyl is selected, wherein R _a and R _{b are selected} from an alkyl and an alkenyl; R ^{5 is selected} from H, alkoxy, morpholinyl, halo, N-alkylpiperazinyl, piperidinyl, pyrrolyl, pyrrolidinyl, pyridyl, -NR _a R _b , acylamino and carbamoyl wherein R _a and R _b may be selected from alkyl and alkenyl; R ⁶ = H; R ^{7 is selected} from H or acylamino. A compound according to claim 4, wherein R ^{5 is selected} from halogen, 4-N-methylpiperazinyl, N-morpholinyl, N-piperidinyl, N-pyrrolyl, N-pyrrolidinyl, N, N-diethylamino, N, N-dimethylmethylamine group and 4 Pyridyl is selected; R ^{3 is selected} from one of the following groups:

wherein X is a halogen; and R ⁷ = H. Compound selected from Compounds 001-032 shown in Table 1. A pharmaceutical composition, the pharmaceutical composition comprising the compound of any of claims 1 to 6 or the pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. Use of the compound according to any one of claims 1 to 6 in the manufacture of drugs for the treatment or prevention of diseases caused by EGFR, BLK, FLT3, HER2, HER4, FLT1, CDK2, JAK2, LCK, LYNA, cKit, PIM1, FGFR3, Or to inhibit EGFR, BLK, FLT3, HER2, HER4, FLT1 , CDK2, JAK2, LCK, LYNA, cKit, PIM1, FGFR3, FGFR1, PDGFRa, PDGFRb, KDR, SRC, ABL, AURB, C-MET, BRAF, PKACa, IKKb, IGF1R, GSK3b, P38a and ERK1. Use according to claim 8, wherein the disease is cancer or an immune disease. Use according to claim 9, wherein the cancer is selected from non-small cell lung cancer, breast cancer, prostate cancer, glial cell tumors, ovarian cancer, head and neck squamous cell carcinoma, cervix cancer, esophageal cancer, liver cancer, kidney cancer, pancreatic cancer, colon cancer, skin cancer, leukemia, lymphoma, gastric cancer, solid tumors, diffuse B-cell lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, follicular lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma, spleen marginal zone lymphoma, plasma cell myeloma, plasma cell tumors, extranodal marginal zone B-cell lymphoma, Marginal zone B-cell lymphoma of the lymph nodes, mantle cell lymphoma, thymic large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma, pain by mycosis fungoides, lymphoblastic lymphoma, T-cell prolymphocytic Leukemia, T-cell granular lymphocytic Le ukemia, aggressive NK cell leukemia, cutaneous T cell lymphoma, plastic large cell lymphoma, peripheral T cell lymphoma, adult T cell lymphoma, acute myeloid leukemia, acute lymphocytic leukemia, acute promyelocytic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic neutrophilic leukemia, acute undifferentiated leukemia, degenerative developmental large cell lymphoma, prolymphocytic leukemia, juvenile myelomonocytic leukemia, adult T-cell ALL-AML mixed myelodysplasia in three cell lines, MLL (mixed lineage leukemia) leukemia, myelodysplastic syndrome, myeloproliferative disorders and multiple myeloma; and the immune disease is selected from arthritis, lupus, inflammatory bowel disease, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, ord thyroiditis, Graves' disease, Felty's syndrome, multiple sclerosis, infectious neuronal inflammation, acute disseminated encephalomyelitis, Addison's disease, aplastic anemia, autoimmune hepatitis, optic neuritis, psoriasis vulgaris, graft-versus-host reaction, transplantation, allergic reaction to transfusion, allergy, type I hypersensitivity, allergic conjunctivitis, allergic rhinitis and atopic dermatitis.